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Secondary sex traits (SSTs) can favour males in intra-sexual competition, allowing females to reliably assess their quality. They can also be connected to other aspects of fitness, such as resistance to parasites and pathogens, as parasites have negative effects on the development of SSTs. Antlers are one of the most recognizable examples of SSTs whose development is regulated by testosterone and reflects the actual condition of the bearer. Elevated testosterone can exaggerate the size of SSTs while impairing the function of the immune system ("The Immunocompetence Handicap Hypothesis") posing a trade-off between antler development and immune function. In this study, we experimentally manipulated the parasite load in captive red deer (Cervus elaphus) males with Ivermectin during antler development for two consecutive years. Expecting an inverse proportionality between parasite load and antler size, we hypothesized the treated deer to have larger antlers than the untreated ones. Our results showed that, following the immunocompetence handicap hypothesis, parasite load was positively associated with testosterone levels. However, the application of Ivermectin suppressed the parasite load of the treated animals but did not lead to the development of larger antlers. Instead, it significantly suppressed the concentration of testosterone in the treated animals, whilst the animals that had higher testosterone also had the highest parasite load. Our findings show that Ivermectin can potentially decrease the levels of testosterone and, consequently, antler size. These findings have important implications for the management of captive populations, especially in contexts where the development of large trophies is desired., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

It is unknown if Leishmania amastigote infections affect hepatocytes and Kupffer cell apoptosis, and the role played by apoptosis in liver lesions in leishmaniasis is still unclear. Clinically affected and subclinically infected dogs with leishmaniosis and uninfected controls were assessed. Parasite load, biochemical markers for evaluation of liver damage, morphometry (area, perimeter, number of inflammatory focus, major and minor diameters), apoptosis in hepatic tissue (hepatocytes, Kupffer cells, and inflammatory infiltrates) and cellularity in inflammatory foci were quantified. The parasite load in clinically affected dogs proved to be higher than in the other groups. All morphometric parameters (area, perimeter, number of inflammatory focus, major and minor diameters) from clinically affected were higher than the values found in the subclinically infected and uninfected control dogs. Only clinically affected dogs presented high levels of ALT, FA, GGT and cholesterol in serum. Strong positive correlation was observed between biochemical markers for evaluation of liver damage (ALT, FA, GGT and cholesterol) and hepatic apoptosis (hepatocytes, Kupffer cells, and inflammation). Clinically affected dogs showed a more intense hepatic lesion. Hepatocytes showed a higher rate of apoptosis in Leishmania-infected dogs than in uninfected control dogs. The Kupffer cell apoptotic index and apoptosis within the inflammatory infiltrates were higher in clinically affected dogs. The apoptotic index evaluated in hepatocytes, Kupffer cells, and inflammatory infiltrates showed a positive correlation with the intensity of the hepatic lesion, parasite load, and clinical status. Apoptotic cells also showed positive immunostaining for TUNEL, Bcl2, and Bax. Our data showed that hepatic apoptosis was related to the severity of liver damage, the progression of infection, and the parasite load in leishmaniasis. Apoptotic regulated cell recruitment modulated the inflammatory response and favored the survival and dissemination of parasites, depending on the clinical status of the Leishmania-infected dogs., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Anilton Cesar Vasconcelos reports financial support and equipment, drugs, or supplies were provided by Minas Gerais State Foundation of Support to the Research., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The aim of this study was to evaluate the histopathological changes to the mammary gland that occur in female dogs with visceral leishmaniosis and to correlate the findings with the parasite load, inflammatory cell profile in mammary tissue and serum progesterone levels. For this, 20 adult female dogs that were naturally infected with Leishmania infantum, not spayed, not pregnant and free from mammary tumors were used. They were divided into two groups: G1 (n = 9) with high serum progesterone levels and G2 (n = 11) with low serum progesterone levels. The parasite load and the immunophenotype of leukocytes infiltrated into the mammary tissue (CD3, CD4, CD8 and MCA874) were evaluated using the immunohistochemical technique. In the mammary gland, chronic inflammatory infiltrate was mainly found in G1, sometimes associated with granulomatous inflammation, higher parasite load and higher density of cells immunolabeled for CD3, CD4, CD8 and MCA874. A significant positive correlation (p < 0.05) was observed between the parasite load and the immunolabeled leukocytes. The influence of the serum progesterone level in the mammary gland of infected female dogs can contribute to the maintenance of an immunosuppressive cell profile and favor the persistence of the parasite in this site., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Ectotherms are vulnerable to environmental changes and their parasites are biological health indicators. Thus, parasite load in ectotherms is expected to show a marked phenology. This study investigates temporal host–parasite dynamics in a lizard community in Eastern Spain during an entire annual activity period. The hosts investigated were Acanthodactylus erythrurus, Psammodromus algirus and Psammodromus edwardsianus, three lizard species coexisting in a mixed habitat of forests and dunes, providing a range of body sizes, ecological requirements and life history traits. Habitat and climate were considered as potential environmental predictors of parasite abundance, while size, body condition and sex as intrinsic predictors. Linear models based on robust estimates were fitted to analyse parasite abundance and prevalence. Ectoparasitic mites and blood parasites from two haemococcidian genera were found: Lankesterella spp. and Schellackia spp. Habitat type was the only predictor explaining the abundance of all parasites, being mostly higher in the forest than in the dunes. The results suggest that particularities in each host–parasite relationship should be accounted even when parasites infect close-related hosts under the same environmental pressures. They also support that lizard parasites can be biomarkers of environmental perturbation, but the relationships need to be carefully interpreted for each host–parasite assemblage.

Background: The association between myocardial parasitic load (MPL) and cardiac biomarkers in Canine Leishmaniasis (CanL) has not been studied., Methods: Dogs with advanced CanL were prospectively recruited and were included if they were euthanised. Prior to euthanasia these variables were assessed: hematocrit, globulin, creatinine, N-terminal-pro brain natriuretic peptide (NT-proBNP), cardiac troponin I (cTnI), blood pressure, urine protein/creatinine ratio and echocardiographic parameters. A left ventricular (LV) sample was taken for histopathology and MPL evaluation by quantitative PCR. Correlation of MPL with all variables was analysed. Dogs with lower and higher histopathology scores were compared., Results: Ten dogs were included. NT-proBNP was 6946 pmol/ (interquartile range [IQR] 3751-9268 pmol/L) and cTnI 4.56 ng/mL (IQR 0.46-13.1 ng/mL). In all dogs, echocardiography showed an increase in LV thickening, and histopathology revealed moderate to severe lympho-plasmocytic myocarditis and/or myocardial cell degeneration. MPL was 215.53 parasites/gram (IQR 21.2-1372.63 parasites/gram). A strong correlation (p < 0.001; R = 0.90; R 2 0.81) with cTnI was observed but correlation with any of the other variables or differences between the two histopathological scores, were not detected., Conclusions: MPL in dogs with advanced CanL shows variable but generally high levels. A strong association between MPL and cTnI was observed, which encourages the exploration of cTnI as a marker in CanL., (© 2021 British Veterinary Association.)

Carotenoid plumage coloration is an important sexually selected trait in many bird species. However, the mechanisms ensuring the honesty of signals based on carotenoid pigments remain unclear. It has recently been suggested that intestinal integrity, which is affected by gut parasites and microbiota and influences nutrient absorption and acquisition, mediates the relationship between carotenoid ornamentation and individual quality. Here, we test whether carotenoid plumage coloration in greenfinches (Chloris chloris) is affected by the treatment of an antibiotic or an antiparasitic drug. We captured wild greenfinches (N = 71) and administered anticoccidial medication toltrazuril (TOLTRA) to one group, antibiotic metronidazole (METRO) to the second group to target trichomonosis, and the third group received no medication. In the METRO group, feathers grown during the experiment had significantly higher chroma of yellow parts, but there was no effect of TOLTRA on feather chroma. The results suggest that METRO increased the efficiency of carotenoid modification or deposition to the feathers rather than nutrient acquisition and/or freed energy resources that could be invested in coloration. Alternatively, though not measured, METRO might have affected microbial community and host physiology as microbial metabolites can modulate mitochondrial and immune function.

Gastrointestinal parasitism is a global problem for grazing ruminants which can be addressed in a sustainable way through breeding animals to be more resistant to disease. This study estimates the genetic parameters of common and new disease phenotypes associated with natural nematode and coccidian infection in Scottish Blackface sheep to underpin future genetic improvement strategies for parasite control. Data on faecal egg counts (FEC) from different species of strongyle parasites and faecal oocyst counts (FOC) from coccidian parasites were collected on 3-month-old lambs together with a faecal soiling score in the breech area dagginess (DAG) and live weight (LWT). Faecal count data were obtained for Strongyles (FEC S ), Nematodirus (FEC N ) and Coccidia (FOC). Data from 3 731 lambs sampled between 2011 and 2017 were included. Faecal egg counts and DAG records were log-transformed prior to analysis. Data were analysed using linear mixed models. Average age at sampling was 92 days with a mean LWT of 24.5 kg. Faecal soiling was not evident in 69% of lambs. Coccidia were the most prevalent parasite (99.5%), while Strongyles and Nematodirus had a prevalence of 95.4% and 72.7%, respectively. Heritability estimates (±SE) were 0.16 ± 0.03, 0.17 ± 0.03, 0.09 ± 0.03, 0.09 ± 0.03 and 0.33 ± 0.04 for FEC S , FEC N , FOC, DAG and LWT, respectively. Strongyles faecal egg count had a strong and positive genetic correlation with FEC N (0.74 ± 0.09) and a moderate positive correlation with FOC (0.39 ± 0.15) while DAG was negatively genetically correlated with LWT (-0.33 ± 0.15). The significant positive genetic correlations between FEC S , FEC N and FOC at 3 months of age show that co-selection of sheep for resistance to these different parasites is feasible. Selection for increased resistance to parasite infection is not expected to adversely affect live BW, as no significant antagonistic genetic correlations were found between LWT and FEC. There were significant antagonistic phenotypic and genetic relationships between DAG and LWT being -0.19 ± 0.02 and -0.33 ± 0.15, respectively, indicating that the expression of the manifestation of disease in lambs may be a more meaningful indicator of the impact of parasite burden on productivity., (Crown Copyright © 2021. Published by Elsevier B.V. All rights reserved.)

The common signs of canine babesiosis caused by an infection with Babesia canis are fever, anorexia, lethargy, pulse alterations, anemia, and occasionally mild icterus. Dogs with these clinical signs can be divided into two groups: those with acute-phase reaction and those with systemic inflammatory response syndrome (SIRS). Factors associated with the occurrence of SIRS in canine babesiosis have not been thoroughly researched. This article outlines a cross-sectional study of 54 client-owned dogs with an acute B. canis infection, and evaluates the differences in age, gender, laboratory findings, parasite load, and seroreactivity against B. canis between the SIRS and the SIRS-free dogs. We have analyzed a complete blood count, serum biochemistry, serum amyloid A, ceruloplasmin, paraoxonase-1, serology, and PCR testing using standard methodologies. The frequency of SIRS among the investigated dogs reached 0.59. Male dogs and those seronegative against B. canis, were more frequent in the SIRS group, whilst age and parasite load could not be associated with the presence of SIRS. Dogs with SIRS had a lower count of total leukocytes, neutrophils, lymphocytes, and monocytes, and a lower concentration of iron and bilirubin compared with SIRS-free dogs. No significant differences in the concentration of acute-phase proteins have been noticed to exist between the groups of dogs. Further, the seronegative dogs had a lower count of lymphocytes and monocytes and a higher parasite load than the seroreactive dogs. Multivariate logistic regression analysis has identified leukopenia (<6 × 10 9 /L) and monocytopenia (<0.2 × 10 9 /L) as independent associates of SIRS in the investigated dogs, thus implying that these routine tests could be used as reliable markers for SIRS., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Reports have shown correlations between the immune response to vector saliva and Leishmaniasis outcome. We followed dogs in an endemic area for two years characterizing resistance or susceptibility to canine visceral leishmaniasis (CVL) according to Leishmania infantum diagnosis and clinical development criteria. Then, we aimed to identify a biosignature based on parasite load, serum biological mediators' interactions, and vector exposure intensity associated with CVL resistance and susceptibility., Methodology/principal Findings: A prospective two-year study was conducted in an area endemic for CVL. Dogs were evaluated at 6-month intervals to determine infection, clinical manifestations, immune profile, and sandfly exposure. CVL resistance or susceptibility was determined upon the conclusion of the study. After two years, 78% of the dogs were infected with L. infantum (53% susceptible and 47% resistant to CVL). Susceptible dogs presented higher splenic parasite load as well as persistence of the parasite during the follow-up, compared to resistant ones. Susceptible dogs also displayed a higher number of correlations among the investigated biological mediators, before and after infection diagnosis. At baseline, anti-saliva antibodies, indicative of exposure to the vector, were detected in 62% of the dogs, reaching 100% in one year. Higher sandfly exposure increased the risk of susceptibility to CVL by 1.6 times (CI: 1.11-2.41). We identified a discriminatory biosignature between the resistant and susceptible dogs assessing splenic parasite load, interaction of biological mediators, PGE2 serum levels and intensity of exposure to sandfly. All these parameters were elevated in susceptible dogs compared to resistant animals., Conclusions/significance: The biosignature identified in our study reinforces the idea that CVL is a complex multifactorial disease that is affected by a set of factors which are correlated and, for a better understanding of CVL, should not be evaluated in an isolated way., Competing Interests: The authors have declared that no competing interests exist.

Leishmaniasis is among the world's most neglected diseases. Dogs are the main reservoirs/hosts of Leishmania infantum, causative agent of both canine and human visceral leishmaniosis. Canine leishmaniasis (CanL) represents a public health problem as one of the most prevalent zoonotic diseases worldwide. Current therapeutics present drawbacks; thus, there is a need for more effective, safer, and cheaper drugs. The aim of this study was to evaluate and to compare the efficacy of oral administration of artesunate or meglumine antimoniate/allopurinol in dogs with clinical leishmaniasis. Forty-two dogs with naturally occurring clinical leishmaniasis were included in this open-label, simple randomized positive-control clinical field trial with 6 months of follow-up. Dogs received meglumine antimoniate 100 mg/kg/day and allopurinol 30 mg/kg/day for 28 days (control group, n = 26) or artesunate 25 mg/kg/day for 6 days (test group, n = 16). The animals were evaluated for their clinical evolution, parasite load (by qPCR) and humoral response at different time points: 0, 30, 90, and 180 days after treatment. Data analyses showed a significant improvement in both groups in clinical scores, parasitemia and antibody titers after treatment. Compared to the control group, the artesunate group showed significantly lower clinical score (P = 0.0001), lower parasitemia (P = 0.0001) and antibody titers after 6 months of follow-up. Compared to baseline values, a rapid, significant reduction (P < 0.012) in antibody levels, 2.28- versus 3.04-fold for the control versus artesunate groups, respectively, was observed 30 days after treatment. Antibody levels continued to decrease further in the artesunate group, where 58% of cases became seronegative at the 6-month follow-up. All qPCR-positive dogs were negative after treatment with artesunate, while 14.3% remained positive with the appearance of two new cases in the control group. Artesunate was well tolerated, and no side effects were recorded. Treatment failures were similar in both groups with 27.27% (6/22), including 18.18% (4/22) mortality in the control group, versus 26.66% (4/15), including 13.33% (2/15) mortality in the artesunate group. This is the first report showing the potential of artesunate in the treatment of dogs with clinical leishmaniasis. Artesunate showed higher efficacy than the current first-line treatment for CanL without any adverse effects. It could be a good alternative chemotherapy for CanL, and may be considered for further studies in human leishmaniases. Further clinical trials are needed to confirm these findings, to determine if there are relapses after treatment and if dogs remain infective to sandflies, to define the ideal therapeutic dosage and duration of treatment with artesunate., Competing Interests: The authors have declared that no competing interests exist.

The tick-borne equine hemoparasite, Theileria equi, is endemic in many parts of the world where prevalence may be high, and most infected horses are apparently healthy but serve as life-long carriers. To determine the factors that affect T. equi dynamics, we followed parasitic loads in apparently healthy horses at four time points during one year. A total of 1094 blood samples were collected from 395 horses, along with ticks and demographic and clinical data. Infection and load of T. equi were tested by PCR and qPCR, and for the spring dataset, infection was also tested serologically by IFAT (n = 268). Theileria equi was molecularly detected in 64.8 % of the horses. The agreement between molecular and serological results was 79.8 % (K > 0.674) and positively correlated with parasitic load. Infection was associated with pale mucus membranes, lower packed cell volume and higher total solids (all P < 0.001), although these changes had only minor clinical importance. While parasitic loads in qPCR-positive samples (n = 561) were generally low (mean = 7.9-10 -4 % parasitized erythrocytes), younger horses showed higher loads, possibly suggesting development of immunity. Infection and parasitic load were associated with housing management and tick exposure, illustrating different patterns of exposure. Endemic stability is suggested in pasture farms with constant exposure to ticks, where parasite prevalence was high (96 %) and associated with T. equi 18S rRNA genotype D, low parasitemia and high antibody titers. Endemic instability can be suggested in case were horses are kept in paddocks (prevalence = 49 %) with intermittent exposure to ticks, where infection was associated with high parasitemia when ticks were present. A steady state is suggested in stabled horses (prevalence = 46 %), with no exposure to ticks; where infection was associated with genotype A, low parasitemia and low antibody titers. The ability to identify different risk groups within endemic areas may improve the administration of suitable treatment and control practices in an effort to reduce the risk of clinical disease., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Parasites co-infecting hosts can interact directly and indirectly to affect parasite growth and disease manifestation. We examined potential interactions between two common parasites of house finches: the bacterium Mycoplasma gallisepticum that causes conjunctivitis and the intestinal coccidian parasite Isospora sp. We quantified coccidia burdens prior to and following experimental infection with M. gallisepticum, exploiting the birds' range of natural coccidia burdens. Birds with greater baseline coccidia burdens developed higher M. gallisepticum loads and longer lasting conjunctivitis following inoculation. However, experimental inoculation with M. gallisepticum did not appear to alter coccidia shedding. Our study suggests that differences in immunocompetence or condition may predispose some finches to more severe infections with both pathogens.

The aim of this study was to evaluate, through qPCR, the prevalence of parasitemia in sick kennel dogs naturally infected by canine leishmaniasis. An evaluation of daily changes of the parasitic load in peripheral blood was also performed. A comprehensive clinical examination and the collection of several samples (blood, lymph node, skin, and conjunctiva) were performed in 140 dogs living in an endemic area. Among these, only the dogs with clinically evident leishmaniasis were enrolled (39/140; 27.9%). Twelve (30.8%) out of 39 showed parasitemia, with a low load (median: 4 Leishmania/ml) despite a high lymph node parasite load (median: 4000 Leishmania/ml) and high IFAT titers (≥ 1:640). Seven sick dogs were sampled every 4 h for 6 times during a 24-h period, in order to obtain light- and dark-span samples. Only one (14.3%) out of the seven serial sampled dogs showed Leishmania DNA in the peripheral blood in two samples (2/42; 4.8%). Surprisingly, Leishmania DNA was also detected in the peripheral blood of asymptomatic dogs, negative to both serology and PCR performed on samples other than blood (6/101; 5.9%). The present study confirms that in canine leishmaniasis parasitemia is uncommon and even transitory. Even if recommended, microscopic examination is confirmed as a low sensitivity method with a lower diagnostic utility in canine leishmaniasis than qPCR. Moreover, circulating Leishmania DNA can be found even in healthy dogs. This finding is important in clinical practice because in endemic areas it suggests a transfusion risk and a possible transmission to the vector.

Background: Amblyomma americanum and Rhipicephalus sanguineus (sensu lato) nymphs commonly feed on and transmit pathogens to dogs (Canis familiaris). Control of immature and adult tick life stages is necessary to fully protect animals. We evaluated efficacy of oral fluralaner (Bravecto ® ) against induced infestations with A. americanum and R. sanguineus (s.l.) nymphs on dogs in two experiments., Methods: In each experiment, 10 dogs were administered oral fluralaner chewable tablets one time on Day 0 at a targeted minimum dose of 25 mg/kg body weight and 10 dogs remained non-treated controls. Dogs were infested with two groups of 50 A. americanum nymphs and two groups of 50 R. sanguineus (s.l.) nymphs on Days -1, 6, 28, 56 and 84. At 48 h and 72 h post-infestation, nymphs were collected from dogs, assessed as live or dead, and enumerated into categories defining attachment and engorgement status. Fluralaner efficacy was determined in separate analyses against all live nymphs and against live-fed nymphs, i.e. live nymphs that were attached to dogs at the time of collection and/or were engorged. Fluralaner was considered effective when mean numbers of live ticks were reduced in fluralaner-treated dogs by ≥ 90%., Results: Fluralaner efficacy against all live and live-fed A. americanum nymphs in the first experiment was > 94% on all collection days. Efficacy against all live R. sanguineus (s.l.) nymphs in the first experiment was  > 96% on all collection days  excluding the 48 h counts for infestations on Days 28 (83.7%), 56 (82.9%) and 84 (86.7%); efficacy against live-fed R. sanguineus (s.l.) nymphs was > 95% on all 48 h/72 h count days. Fluralaner efficacy against all live A. americanum nymphs in the second experiment was > 93% on all collection days for 8 weeks excluding the 48 h count for infestation on Day 56 (87.8%); efficacy against live-fed A. americanum nymphs was > 91% on all count days for 8 weeks. Efficacy against all live R. sanguineus (s.l.) nymphs in the  second experiment was > 91% on all 72 h collection days  except for infestations on Days 28 (76.8%) and 56 (86.3%); efficacy against live-fed R. sanguineus (s.l.) nymphs was 100% on all 72 h count days., Conclusions: A single administration of oral fluralaner to dogs is effective against A. americanum and R. sanguineus (s.l.) nymphs for up to 12 weeks.

Parasitic infections are associated with profound changes in the structure and function of the gut microbiome in various host-parasite systems. Here we examined the microbial composition and function in the abomasum, proximal colon and feces of Haemonchus contortus-infected goats after a partial anthelmintic drug clearance. A single-dose treatment of H. contortus-infected goats with Cydectin (moxidectin) resulted in an 83.9 % and 61.8 % reduction in fecal egg counts (EPG) and worm burden, respectively (P < 0.01), and restored abomasal pH to a normal baseline level. The treatment significantly increased the abundance of Proteobacteria, particularly that of Campylobacter, in the proximal colon. It also significantly affected several basic pathways, including bacterial secretion, butyrate metabolism, and LPS biosynthesis, and seemingly reduced the cellulolytic capacity in the colon. Several network modules displayed a strong correlation with EPG and worm burden. The Mantel test indicated a strong correlation between treatment related network topologies of the operational taxonomic units (OTU) belonging to Actinobacteria and Rikenellaceae and EPG and worm burden levels, respectively. Furthermore, microbial signatures that may better predict anthelmintic efficacy were identified. A signature or balance represented by the log ratio of the abundance of Verrucomicrobiaceae and Camplyobacteraceae had a strong correlation with EPG (r = 0.80). These novel insights into the interactions between H. contortus and gut microbiome in the caprine host and the consequence of a partial anthelmintic clearance on animal health and well-being may facilitate the design of more effective next-generation anthelmintics., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Allopreening occurs when 1 bird preens another bird. The behavior is normally directed at the head and neck of the recipient, i.e., regions that the bird cannot self-preen. Studies of penguins, pigeons, and other groups of birds suggest that allopreening plays a role in the control of ectoparasites, such as ticks and feather lice. However, it is not known whether allopreening increases in response to increases in parasite load, or whether it is a programmed response that occurs independently of parasite load. We conducted a laboratory experiment using wild-caught rock pigeons ( Columba livia ) to test the relationship between ectoparasite load and allopreening rate. We added feather lice ( Columbicola columbae ) to captive pigeons and tested for changes in allopreening rates compared to control birds with no lice added. Allopreening rates did not change in response to the addition of lice. Interestingly, however, our data revealed a negative correlation between allopreening and self-preening rates.

An individual-based model was parameterized to explore the impact of a crustacean ectoparasite (sea louse, Lepeophtheirus salmonis & Caligus spp.) on migrating Atlantic salmon smolt. The model explores how environmental and intrinsic factors can modulate the effect of sea lice on survival, growth and maturation of Atlantic salmon at sea. Relative to other effects, the parasite infestation pressure from fish farms and the encounter process emerge as the most important parameters. Although small variations in parasite-induced mortality may be masked by variable environmental effects, episodes of high infestation pressure from fish farms should be observable in wild populations of Atlantic salmon if laboratory studies accurately reflect the physiological effects of sea lice. Increases in temperature in the model negatively influenced fish survival by affecting the development time of the parasite at a rate that was not compensated for by the growth of the host. Discharge from rivers was parameterized to increase migration speed and influenced parasite induced mortality by decreasing time spent in areas with increased infestation pressure. Initial size and growth of the host was inversely related to the impact of the parasite because of size-dependent parasite-induced mortality in the early phase of migration. Overall, the model illustrates how environmental factors modulate effects on the host population by impacting either the parasite load or the relative effect of the parasite. The results suggest that linking population-level effects to parasite infestation pressure across climatic and environmental gradients may be challenging without correctly accounting for these effects.

Background: Toxoplasmosis is a zoonotic parasitic disease caused by Toxoplasma gondii, a widespread protozoan in the phylum Apicomplexa. In Europe, several studies have demonstrated the presence of the parasite in tissues of wild boars (Sus scrofa), but no data exists on the T. gondii load in tissues which in turn may be an useful way to assess the infection risk for the consumer of wild boar meat., Methods: We sampled and tested a total of 472 tissue samples of brain, heart and masseter muscle from 177 wild boars from the Campania region of southern Italy by real-time PCR analyses for detection and quantification of T. gondii. The sensitivity and specificity of the method were calculated by ROC analysis curves., Results: PCR analysis revealed the presence of T. gondii in tissue samples of 78 out of 177 (44%) wild boars. In general, the brain presented the highest PCR prevalence (31%), followed by the heart (28.3%) and the masseter muscle (24.2%), with the highest estimated parasite numbers observed in the brain followed by the heart and masseter muscle. The PCR method showed an excellent discriminating ability for each of the examined tissues. According to the ROC analysis curves, the respective sensitivity and specificity were 99 and 100% for masseter muscle, 98 and 98% for brain and 96 and 98% for heart samples., Conclusions: The high prevalence of infection here detected suggests a widespread distribution of the parasite in the wildlife of the Campania region of southern Italy. The T. gondii burdens detected may potentially represent a source of infection for humans.

Background: Visceral leishmaniasis (VL) is an infectious disease with a variety of clinical signs. The main form of parasite transmission to humans and other mammalian hosts is through the bite of infected arthropod females with Lutzomyia longipalpis serving as the main vector in the Americas. Dogs are the main urban domestic reservoirs of the parasite and the main source of vector infection due to their high prevalence in endemic areas and the large number of parasites in the skin of infected animals. Although miltefosine has been used in Europe since 2002 for treatment of VL infected dogs, in the Americas the treatment of dogs has not been recommended. Therefore, this study aimed to evaluate efficacy of miltefosine observing a reduction of clinical signs in infected dogs and the infectiveness to the vector by Leishmania (L.) infantum., Methods: To our knowledge, this is the first controlled study using qPCR and xenodiagnosis to evaluate the efficacy of miltefosine (Milteforan®, Virbac) as a single treatment in Brazil. Thirty-five adult dogs with canine visceral leishmaniasis (CVL), confirmed by clinical and laboratory tests, were included in this study. They received miltefosine at a dose of 2 mg/kg every 24 h for 28 days. The dogs were observed over a three-month period, during which clinical evaluations based on a scoring system were conducted at pre-established times. Parasite load was assessed by cytology and real-time polymerase chain reaction (qPCR). Transmissibility to the vector was evaluated by xenodiagnosis., Results: At the end of the period, the following were observed: (i) the remission of clinical signs with a reduction in clinical scores for 94.2% of the animals; (ii) a statistically significant reduction (98.7%) in parasitic load by qPCR; and (iii) a reduction in infectivity to sand flies. After treatment, 74.2% of the animals remained or had become non-infectious., Conclusions: Our study indicates that the use of miltefosine administered orally for 4 weeks contributes to a clinical improvement and reduction in infectivity of dogs to L. infantum.

Haemonchus contortus is a nematode parasite that establishes in the abomasum of ruminants, it has a cosmopolitan distribution and is a worldwide health problem for livestock. As a first line of defence against parasites, behaviour can help to prevent or fight infection, and may even serve as a method of early presumptive diagnosis. Parasites can affect performance of cattle and cause significant economic losses. The aim of this study was to determine the behavioural and productive changes induced by an experimental infection with H. contortus L3 in bovines. We used 32 dewormed bull calves, randomly divided into two groups, 8 no inoculated controls and 24 that were inoculated with 4000 L3 of H. contortus. Inoculation did not influence haematocrit or haemoglobin values at 0, 28 and 42 days post infection (P> 0.20); however, an increase in the frequency of urination (P = 0.0001) and defecation (P = 0.0001), number of steps (P < 0.001) and self-grooming (P < 0.01) events were observed, even in inoculated animals in which not parasite eggs were found in faeces. During the first 28 days post-inoculation with H. contortus, feedlot performance was not affected (P = 0.16), but during the last 14 days (29-42) inoculated animals gained 15% less weight compared to controls (P = 0.04). Over 42 days, inoculated calves showed a 28% poorer feed efficiency during the 42 days post-inoculation than controls (P < 0.01). Across the experiment, calves inoculated with H. contortus obtained less net energy both for maintenance as for gain from diet (P < 0.01). It is concluded that both, performance and behaviour were modified in feedlot bull-calves parasitized by H. contortus, supporting the practice of deworming when eggs of this parasite are detected in the faeces even in low amounts., (Copyright © 2018. Published by Elsevier Inc.)

Enteromyxum leei has been reported to cause emaciation disease in various fish species. To determine the effect of parasite intensity on cultured olive flounder Paralichthys olivaceus, we investigated the relationship between the relative condition factor (rCF = CF/standard CF × 100) and parasite load with quantitative polymerase chain reaction (qPCR) and the challenge test. A total of 57 cultured olive flounders were obtained from 11 fish farms and divided into five groups based on their rCF. We investigated the parasite intensity in the posterior intestine of the fish. The parasite load was closely matched to severe loss of body weight. In addition, olive flounders were inoculated either orally or anally with intestinal scrapings of infected fish or phosphate-buffered saline. The fish were reared at natural water temperature and transferred to different tanks, and the water temperature was adjusted to 20°C after 6 weeks of inoculation. When the water temperature was increased to 20°C, the rCF decreased in the experimentally infected group. The results demonstrated that qPCR can be utilized to determine the relative abundance of E. leei in olive flounders and water temperature is an important factor to track the progress of the emaciation disease., (© 2018 John Wiley & Sons Ltd.)

The effects of introgression on parasitism in brook charr Salvelinus fontinalis were investigated in 28 lakes with various levels of stocking in Québec, Canada. No effect of genetic background on parasitism was found at the individual level. Body length seemed to explain most of the variation observed at this level, with largest fish being more infected. However, lakes with the greater average domestic genetic background were found to display significantly lower parasite prevalence and diversity. Since our results indicate no effect of domestic genes at the individual level, the negative association with introgression found at the population level may be mainly attributed to differences in intrinsic environmental quality of lakes (e.g. fishing pressure, availability of food resources, abiotic characteristics)., (© 2018 The Fisheries Society of the British Isles.)

Eosinophils are traditionally associated with the immune response against helminth parasites. However, several studies have demonstrated that these cells have a role regarding protective immunity in leishmaniasis. Here, we examined the relationship between the presence of eosinophils and parasite load in biopsy samples from dogs, obtained through fine needle puncture and aspiration of lymph nodes. Histological slides containing biopsy material from lymph nodes of dogs with canine visceral leishmaniasis and healthy dogs were used to obtain baseline eosinophil counts. Subsequently, scrapings were taken from slides for DNA extraction and determination of parasite load, using real-time PCR (qRT-PCR). Additionally, production of nitric oxide (NO) and reactive oxygen species (ROS) levels by eosinophils in the peripheral blood of dogs with canine visceral leishmaniasis and healthy dogs was measured. The eosinophil percentage were higher in lymph nodes of infected group, and the parasite load showed a significant negative correlation with the eosinophil count. The production of NO and ROS by eosinophils in the peripheral blood was higher in the dogs with canine visceral leishmaniasis. All the results together suggest that eosinophils may participate in antileishmanial immunity in canine visceral leishmaniasis., (© 2018 John Wiley & Sons Ltd.)

Classical research on animal toxicity has focused on the role of toxins in protection against predators, but recent studies suggest these same compounds can offer a powerful defense against parasites and infectious diseases. Newts in the genus Taricha are brightly coloured and contain the potent neurotoxin, tetrodotoxin (TTX), which is hypothesized to have evolved as a defense against vertebrate predators such as garter snakes. However, newt populations often vary dramatically in toxicity, which is only partially explained by predation pressure. The primary aim of this study was to evaluate the relationships between TTX concentration and infection by parasites. By systematically assessing micro- and macroparasite infections among 345 adult newts (sympatric populations of Taricha granulosa and T. torosa), we detected 18 unique taxa of helminths, fungi, viruses and protozoans. For both newt species, per-host concentrations of TTX, which varied from undetectable to >60 μg/cm 2 skin, negatively predicted overall parasite richness as well as the likelihood of infection by the chytrid fungus, Batrachochytrium dendrobatidis, and ranavirus. No such effect was found on infection load among infected hosts. Despite commonly occurring at the same wetlands, T. torosa supported higher parasite richness and average infection load than T. granulosa. Host body size and sex (females > males) tended to positively predict infection levels in both species. For hosts in which we quantified leucocyte profiles, total white blood cell count correlated positively with both parasite richness and total infection load. By coupling data on host toxicity and infection by a broad range of micro- and macroparasites, these results suggest that-alongside its effects on predators-tetrodotoxin may help protect newts against parasitic infections, highlighting the importance of integrative research on animal chemistry, immunological defenses and natural enemy ecology., (© 2018 The Authors. Journal of Animal Ecology © 2018 British Ecological Society.)

The most commonly used culture medium for the in vitro isolation of Leishmania spp. from canine biological samples is biphasic Novy-MacNeal-Nicolle (NNN) medium, whose solid phase is prepared using rabbit blood. Leishmania infantum parasites from natural infections are highly sensitive and demanding for growth in axenic conditions when firstly obtained from the dog's body. The objective of this study was to evaluate whether NNN medium (NNN-test) prepared with chicken blood (NNN-C), ox blood (NNN-O), horse blood (NNN-H) or sheep blood (NNN-S) was viable for the isolation of parasites from naturally infected dogs, in an endemic area for visceral leishmaniasis caused by L. infantum. Spleen aspirates from six dogs previously diagnosed as infected by parasitological methods were simultaneously inoculated in each NNN-test medium, including the conventional medium prepared with rabbit blood (NNN-R), and the cultures were examined for three weeks under optic microscopy. Spleen samples were also analyzed for parasite loads by quantitative PCR (qPCR). Cultures from three of the six dogs (50%) were positive in at least one of the NNN-test media: one sample presented the highest spleen parasite load by qPCR (1.19 × 10 4 parasites/mL) and was positive in all test media; the second sample presented parasitic isolation in the first week of culture in all inoculated media, of which the NNN-C medium had the highest mean parasite count (NNN-C = 23.5 × 10 4 /mL vs. NNN-R = 3.25 × 10 4 /mL); the third sample was positive only in the NNN-S medium besides the conventional control NNN-R. Cultures from the three remaining dogs were negative in all NNN media, including the control and test media; of those three dogs, two presented the lowest spleen parasitic loads according to qPCR. Blood from chicken, ox, horse and sheep shown to be viable for the preparation of NNN culture medium for the primary isolation of L. infantum from samples of naturally infected dogs and can be considered as an alternative to rabbit blood when necessary., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objectives: To quantify (by qPCR) the density of Demodex canis mites in the skin of dogs with demodicosis and in healthy dogs, as well as measuring the serum concentrations of interleukin (IL)-1β, IL-6, IL-8, IL-10, IL-12, and tumour necrosis factor-alfa (TNF-α)., Methods: Fifty-four dogs were divided into three groups: localized demodicosis (LD, n = 16), generalized demodicosis (GD, n = 22), and control group (CG, n = 16). All dogs were subjected to skin scraping, blood collection, and skin biopsy. DNA extraction was performed and the parasite density was established by qPCR. Serum cytokine concentrations were obtained by flow cytometry., Results: The median number of mites in the skin of the GD (6.2 × 10 4 copies/μL) and LD dogs (1.2 × 10 4 copies/μL) was statistically higher than that in the CG dogs (8.7 × 10 2 copies/μL). Whereas there were no significant differences in median IL-1β, IL-8, IL-10, IL-12, and TNF-α levels among the study groups, there was a statistically higher IL-6 concentration in the LD dogs than in the healthy dogs., Conclusions: According to our results, qPCR is an effective method for measuring the density of D. canis in the canine integument. In addition, the activation of the acute-phase immune response in localized demodicosis can be induced by IL-6 activity., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Captive-bred animals, widely used in reintroduction programmes, are often immunologically naïve and more susceptible to pathogens. We analysed infection of invasive blood-sucking nematode Ashworthius sidemi in captive-bred European bison (Bison bonasus) released to the wild in the Białowieża Forest (Poland). Mean A. sidemi infection intensity of released bison (29,137 nematodes) was over threefold higher than in wild bison (8756). It indicates a rapid acquisition and increase in the infection intensity in previously dewormed bison released from captivity. Thus, reintroduction programmes should consider the impact of pathogens and involve controlled exposure of captive animals to specific parasites prior to release.

Among the topics related to invasion science, the least studied are parasite co-introduction and spillback. This leads to an uncertainty in invasion ecology theories and applications to management. Therefore, the present study brings a systematic review of published information on the metazoan parasite fauna of Micropterus salmoides, a widely introduced fish, with the aim of comparing information about the composition and richness of the associated parasite communities in its native and introduced regions. This review demonstrates that there were twice as many studies of M. salmoides in its native region in comparison with introduced regions, although most of the studies focused on the analysis of a single species or taxon of parasite. This bias impacts the number of parasite species observed and, consequently, the apparent importance of enemy release in introduced regions. The composition of the parasite community in the two regions showed high similarity, which indicates the introduction and acquisition of parasites in introduced regions. Otherwise, there was no pattern related to the geographic distance, highlighting the influence of the propagule pressure and vector strength on the introduction of novel parasites. This illustrates the importance of vector strength on fish-parasite co-introduction and the necessity of new research examining host-parasite interactions with the parasite community of the invaded ecosystems. We still do not know the major influences of the composition of the parasite fauna of M. salmoides or how we can manage to develop a more restrictive vector pathway of introduction. The future of our ecosystems depends on how to account for current and future interactions among novel interactions, habitat, and climate change.

In this paper, we have described for the first time a semiquantitative method to evaluate histopathological damage, taking the degree of Neoechinorhynchus buttnerae attachment to the intestinal wall of the tambaqui (Colossoma macropomum), an important species in Brazilian aquaculture, into account. Twelve specimens of tambaqui were collected from a fish farm. Their bowels were removed and divided into seven morphologically distinct portions according to density and distribution of the parasite studies. Fragments from each fraction were histologically processed and analyzed. There was a clear preference on the part of N. buttnerae for the intermediate regions of the intestinal tube, where the highest densities were recorded. The intensity of damage to the host, estimated by calculating the Histopathological Alteration Index (HAI), showed severe and irreversible changes only where the parasite had its proboscis penetrated into the intestine wall.

Northern bobwhite quail (Colinus virginianus) are a highly sought-after game bird in the Rolling Plains of West Texas. Unfortunately, bobwhite populations in this area are subject to dramatic fluctuations and have been steadily decreasing over the past several decades. While many factors have been investigated as potential mechanisms of cyclic and declining bobwhite numbers, the effect of parasites on bobwhite populations has historically been undervalued. Between December 2017 and February 2018, we received 21 hunter-shot bobwhite from Garza and Mitchell counties in Texas and found peak caecal worm (Aulonocephalus pennula) and eyeworm (Oxyspirura petrowi) burdens averaging 599 and 44, respectively. These represent the highest average parasite loads we have documented in bobwhite from the Rolling Plains thus far and are coincident with widespread reports of declining bobwhite abundance. These elevated infections also followed a high point in bobwhite populations in the Rolling Plains, and our observations of infection dynamics during this time reflect other instances of potential parasite-induced host mortality. While the sample discussed in this communication is small, our findings highlight the need for additional research into how parasites may affect bobwhite population fluctuations in this region.

Parasites have profound fitness effects on their hosts, yet these are often sub-lethal, making them difficult to understand and quantify. A principal sub-lethal mechanism that reduces fitness is parasite-induced increase in energetic costs of specific behaviours, potentially resulting in changes to time and energy budgets. However, quantifying the influence of parasites on these costs has not been undertaken in free-living animals. We used accelerometers to estimate energy expenditure on flying, diving and resting, in relation to a natural gradient of endo-parasite loads in a wild population of European shags Phalacrocorax aristotelis We found that flight costs were 10% higher in adult females with higher parasite loads and these individuals spent 44% less time flying than females with lower parasite loads. There was no evidence for an effect of parasite load on daily energy expenditure, suggesting the existence of an energy ceiling, with the increase in cost of flight compensated for by a reduction in flight duration. These behaviour specific costs of parasitism will have knock-on effects on reproductive success, if constraints on foraging behaviour detrimentally affect provisioning of young. The findings emphasize the importance of natural parasite loads in shaping the ecology and life-history of their hosts, which can have significant population level consequences., (© 2018 The Authors.)

Experimental infections in pregnant sheep have been focused on studying the effect of the time of challenge on the outcome of N. caninum infection, whereas the impact of the dose and route of challenge has not been studied in depth. Therefore, clinical outcome, immune responses, parasite detection and burden, and lesion severity in placental tissues and foetal brains were investigated in 90-day-pregnant sheep inoculated intravenously with 10 5 (G1), 10 4 (G2), 10 3 (G3), or 10 2 (G4) tachyzoites or subcutaneously with 10 4 (G5) tachyzoites of the virulent Nc-Spain7 isolate and an uninfected group (G6). Comparing challenge doses, G1 was the only group that had 100% abortion. Likewise, IFNγ levels in G1 increased earlier than those in other intravenously infected groups, and IgG levels on day 21 post-infection (pi) were higher in G1 than those in other intravenously infected groups. Concerning vertical transmission, G1 shows a higher parasite burden in the foetal brain than did G2 and G3. Comparing routes of administration, no differences in foetal survival rate or parasite load in the foetal brain were found. Although G2 had higher IFNγ levels than G5 on day 10 pi, no differences were found in humoral immune responses. Because the outcome after intravenous infection with 10 5 tachyzoites was similar to that observed after intravenous infection with 10 6 tachyzoites used in a previous work (100% abortion and vertical transmission), we conclude that it may be reasonable to use 10 5 tachyzoites administered by the intravenous route in further experiments when assessing drugs or vaccine candidates.

A key goal in the control of canine visceral leishmaniosis (CVL) has been the development of vaccines with a highly protective capability to interrupt the parasite transmission cycle. However, in addition to promising vaccine searches, researchers have sought to develop new drugs capable of eliminating parasites in humans and dogs. With that in mind, this study analyzed an immunotherapeutic approach in dogs naturally infected with Leishmania infantum. Fourteen dogs were divided into two groups and received a protocol of immunotherapeutic treatment with five doses of total antigens of Leishmania amazonensis or total antigens of L. amazonensis plus saponin (LaSap). All the animals were evaluated before and 90 and 180 days after treatment, hematology, liver and renal biochemical analyzes, serology, lymphoproliferation, and parasite load by qPCR. The results of immunotherapy with the LaSap vaccine were promising since it was able to preserve hematological and biochemical parameters, as well as improve the clinical status, reduce serum levels of IgG, induce a lymphoproliferative capacity against soluble antigens of L. infantum, and provide a marked reduction in the parasite load after LaSap immunotherapeutic treatment. The immunotherapy data demonstrated that LaSap offered the best formulation to induce clinical cure associated with a parasite load reduction in the skin. However, after 180 days of treatment, the animals again showed a slight increase in parasitism, indicating that immunotherapy does not promote sterilizing cure and a new immunotherapeutic intervention would be necessary to maintain low parasitism in dogs., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The spleen is one of the main affected organs in canine visceral leishmaniasis (CVL). Disorganization of the splenic white pulp (SWP) has been associated with immunosuppression and disease progression. This study aims to assess structural and cellular changes in the splenic extracellular matrix of dogs with CVL, correlating these changes with the parasite load and clinical signs. Splenic fragments were collected from 41 naturally infected animals for parasite load quantification by quantitative PCR, histopathological analysis and immunohistochemistry for CD3+, CD4+, and CD8+ T cells; CD21+ B cells; Ki-67+, IFN-γ+, and IL-10+ cells; and the MMP-9 and ADAM-10 enzymes. Laminin, collagen and fibronectin deposition were also evaluated. The animals were grouped according to the level of SWP organization. SWP disorganization was accompanied by a reduction in the quantity of lymphoid follicles/mm2 (p > 0.0001). Animals with moderate to intense SWP disorganization showed more clinical signs (p = 0.021), higher laminin (p = 0.045) and collagen deposition (p = 0.036), higher MMP-9 expression (p = 0.035) and lower numbers of CD4+ T cells (p = 0.027) in the spleen than the animals with organized SWP. These data suggest that splenic structure and function are drastically altered and compromised during CVL.

In most host-parasite systems, variation in parasite burden among hosts drives transmission dynamics. Heavily infected individuals introduce disproportionate numbers of infective stages into host populations or surrounding environments, causing sharp increases in frequency of infection. Parasite aggregation within host populations may result from variation among hosts in exposure to infective propagules and probability of subsequent establishment of parasites in the host. This is because individual host heterogeneities contribute to a pattern of parasite overdispersion that emerges at the population level. We quantified relative roles of host exposure and parasite establishment in producing variation in parasite burdens, to predict which hosts are more likely to bear heavy burdens, using big brown bats (Eptesicus fuscus) and their helminths as a model system. We captured bats from seven colonies in Michigan and Indiana, USA, assessed their helminth burdens, and collected data on intrinsic and extrinsic variables related to exposure, establishment, or both. Digenetic trematodes had the highest prevalence and mean abundance while cestodes and nematodes had much lower prevalence and mean abundance. Structural equation modeling revealed that best-fitting models to explain variations in parasite burden included genetic heterozygosity and immunocompetence as well as distance to the nearest water source and the year of host capture. Thus, both differential host exposure and differential parasite establishment significantly influence heterogeneous helminth burdens, thus driving population-level patterns of parasite aggregation., (Copyright © 2018 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Atlantic salmon populations across the world have diverse ecological and evolutionary histories, from wild anadromous or landlocked, to domestication and genetic modification. The natural host behaviours confer protection from infestation by ectoparasitic salmon lice Lepeophtheirus salmonis, yet whether genetic origin results in different behaviours and thus susceptibility to infestation is unknown. In common garden experiments, we tested antiparasite behaviours, susceptibility and retention of salmon lice in wild anadromous, wild landlocked, domesticated and genetically modified domesticated strains. Within domesticated strains, we tested two infestation histories (previously infested and naïve) and a new phenotype (albino colouring). Farmed stocks initially acquired 24%-44% higher levels of parasite density than the wild and landlocked strains. Burst swimming and displacement behaviours were higher in the domesticated groups, and jumping was more prevalent in the domesticated strains. At 34 days post-infestation, domesticated strains and the wild anadromous strain did not differ significantly from each other; however, landlocked salmon had increased infestation levels considerably. Domesticated strains lost ~20% (±9.9%-16.5%; 95% CI) of their initial parasite load, while parasite load increased by 5.5% (±30.1%) for wild salmon and 20.1% (±28.5%) in landlocked salmon. This study provides early evidence for diverged host-parasite interactions associated with domestication in this system., (© 2017 John Wiley & Sons Ltd.)

Pathogen-induced host phenotypic changes are widespread phenomena that can dramatically influence host-vector interactions. Enhanced vector attraction to infected hosts has been reported in a variety of host-pathogen systems, and has given rise to the parasite manipulation hypothesis whereby pathogens may adaptively modify host phenotypes to increase transmission from host to host. However, host phenotypic changes do not always favour the transmission of pathogens, as random host choice, reduced host attractiveness and even host avoidance after infection have also been reported. Thus, the effects of hosts' parasitic infections on vector feeding behaviour and on the likelihood of parasite transmission remain unclear. Here, we experimentally tested how host infection status and infection intensity with avian Plasmodium affect mosquito feeding patterns in house sparrows (Passer domesticus). In separate experiments, mosquitoes were allowed to bite pairs containing (i) one infected and one uninfected bird and (ii) two infected birds, one of which treated with the antimalarial drug, primaquine. We found that mosquitoes fed randomly when exposed to both infected and uninfected birds. However, when mosquitoes were exposed only to infected individuals, they preferred to bite the non-treated birds. These results suggest that the malarial parasite load rather than the infection itself plays a key role in mosquito attraction. Our findings partially support the parasite manipulation hypothesis, which probably operates via a reduction in defensive behaviour, and highlights the importance of considering parasite load in studies on host-vector-pathogen interactions., (Copyright © 2017 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Leishmania infantum causes from subclinical infection to severe disease in humans and dogs. The spleen is one of the organs most affected by the infection. Although evidence exists that the parasitic load distribution and histological alterations may not be homogeneous in the affected organs of naturally infected individuals, it has not been formally demonstrated using the current techniques used for studying the disease. In six dogs naturally infected with Leishmania, parasitic load and histological changes were compared in samples collected from the lower, middle and upper third of the spleen. Parasitic load in the spleen of the group of dogs was variable, revealing a difference of 61 times between animals with the lowest and the highest parasitism. The set of parasitic load values of each dog showed a cluster trend, when compared to the other animals. Nevertheless, the parasitic load values of each dog showed a variation ranging from 3.2 to 34.7 times between lowest and highest value. Histological changes showed recognizable variation in frequency (granulomas) or intensity (perisplenitis) in the spleen of 2 out of the 6 dogs. The agreement of histological findings between samples collected from the different thirds of the spleen was good (kappa coeficient, 0.61-0.80) very good (0.81-0.99) or perfect (1.00), for most of the parameters analyzed. Variability of parasitic load and, to a lesser extent, histological changes in spleen of dogs with visceral leishmaniasis is observed. Such variability may be taken in account in the design of studies on pathogenesis, vaccine and therapeutic drug development., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The Schlei fjord in northern Germany is the recipient water of a comprehensive eel, Anguilla anguilla (L.), stocking programme. Since 2015, stocked eels become alizarin red S marked, but to date no control mechanism is implemented in this stock enhancement measure to prevent anthropogenic spreading of diseases. Consequentially, it was possible that farmed stocking cohorts of 2015 and 2016 (in total ca. 1040 kg) were subsequently tested positive for anguillid herpesvirus 1 (AngHV 1). For this study, 100 eels [total length (TL) 24.3-72.9 cm, age ca. 1-6 years] were caught in 2016 and investigated with regard to AngHV 1 infection, parasite load (Anguillicoloides crassus) and body conditions. 68% of the eels were found to be virus positive while larger specimens were more often infected. In addition, a fitted generalized linear model (area under the curve = 0.741) demonstrated that an increase in individual TL is accompanied with an increased risk of clinically relevant virus loads. Anguillicoloides crassus turned out to be an important stressor for eels, because parasite and virus load revealed a significant positive correlation. The results of this study evidently show the urgent need of a disease containment strategy for eel stocking programmes., (© 2017 John Wiley & Sons Ltd.)

First-line treatment for canine leishmaniosis (CanL) is N-methylglucamine antimoniate (MGA) combined with allopurinol. However, in some dogs allopurinol may induce hyperxanthinuria leading to urolithiasis. Moreover, allopurinol resistance has recently been described in Leishmania infantum isolates from treated dogs with a relapse of the disease. Alternative treatments are thus needed. Since the type of host immune response strongly influences CanL progression and prognosis, dogs could benefit from treatments targeted at modulating such response, such as nucleotides and active hexose correlated compound (AHCC). The aim of this study was to evaluate the effects of an oral combination of nucleotides and AHCC in dogs with clinical leishmaniosis. Sixty-nine dogs with naturally-occurring clinical leishmaniosis were included in this multicenter, open-label, positively-controlled clinical trial and randomized to receive 10mg/kg allopurinol PO BID (allopurinol group) or 17mg/kg AHCC plus 32mg/kg nucleotides PO SID (supplement group) for 180 days. All dogs were also given 50mg/kg MGA SC BID during the first 28 days. At the time points 0, 30, and 180 days of the trial, dogs underwent a clinical examination, and blood, urine, and bone marrow samples were submitted for analytical tests. Final data analyses (allopurinol group: n=29; supplement group: n=24) revealed a significant improvement in both groups in clinical scores and ELISA-determined antibody titers after treatment. However, the supplement group showed a significantly lower clinical score (P=0.005) and significantly higher antibody titers (P=0.032) after 180 days, compared to the allopurinol group. RT-PCR parasite loads were reduced in groups (mean±SD supplement: 0.38±0.56 vs 5.23±18.9; allopurinol: 0.45±1.47 vs 3.09±8.36 parasites/ng of DNA), but there were no significant differences over time or between groups. During the study, 12 dogs in the allopurinol group developed xanthinuria (41%) compared to no dogs (0%) in the supplement group (P=0.000). Both treatments led to significantly increased CD4+/CD8+ ratio, and improvements in protein electrophoretic pattern and acute phase response. In conclusion, 6-month oral treatment with nucleotides and AHCC in addition to MGA showed similar efficacy to the current first-line treatment for CanL, without producing xanthinuria. This combination could be a good alternative to MGA-allopurinol combination treatment for CanL, especially for dogs suffering allopurinol-related adverse events., (Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.)

Microparasites play an important role in the demography, ecology and evolution of Pacific salmonids. As salmon stocks continue to decline and the impacts of global climate change on fish populations become apparent, a greater understanding of microparasites in wild salmon populations is warranted. We used high-throughput, quantitative PCR (HT-qRT-PCR) to rapidly screen 82 adult Chinook salmon from five geographically or genetically distinct groups (mostly returning to tributaries of the Fraser River) for 45 microparasite taxa. We detected 20 microparasite species, four of which have not previously been documented in Chinook salmon, and four of which have not been previously detected in any salmonids in the Fraser River. Comparisons of microparasite load to blood plasma variables revealed some positive associations between Flavobacterium psychrophilum, Cryptobia salmositica and Ceratonova shasta and physiological indices suggestive of morbidity. We include a comparison of our findings for each microparasite taxa with previous knowledge of its distribution in British Columbia., (© 2017 John Wiley & Sons Ltd.)

Background: Normal-looking skin of dogs with leishmaniosis frequently shows microscopic lesions along with the presence of Leishmania amastigotes. However, histological lesions with or without detection of amastigotes might not occur in less severe clinical cases. In addition, comparative studies between paired clinically-lesioned and normal-looking skin samples from dogs with different disease severity are lacking. The objective of this study was to compare histological and parasitological findings by Leishmania immunohistochemistry (IHC) and quantitative PCR (qPCR) on paired clinically-lesioned and normal-looking skin biopsies from 25 dogs with different clinical stages of leishmaniosis, 11 with stage I-mild disease (papular dermatitis) and 14 with stage II-III (ulcerative or exfoliative dermatitis)., Results: The study demonstrated microscopic lesions in 14 out of 25 (56%) samples from normal-looking skin biopsies. In those samples, perivascular to interstitial dermatitis composed by macrophages with lymphocytes and plasma cells was observed mainly in the superficial and mid-dermis. The intensity of the dermatitis was mild to moderate and always less prominent than in the clinically-lesioned skin. In normal-looking skin samples, the presence of parasites was detected by histology, IHC and qPCR in 5/25 (20%), 8/25 (32%) and 18/25 (72%), respectively. Leishmania was encountered in 11/25 (44%), 23/25 (92%) and 25/25 (100%) of clinically-lesioned skin samples by histology, IHC and qPCR, respectively. Normal-looking skin from dogs with stage I-mild disease was less frequently inflamed (P = 0.0172). Furthermore, Leishmania was more easily demonstrated by histology (P = 0.0464), IHC (P = 0.0421) or qPCR (P = 0.0068) in normal-looking skin of dogs with stage II-III-moderate to severe disease. In addition, in the latter group, there was a significantly higher parasite load studied by means of qPCR than in dogs with less severe disease (P = 0.043). Clinically-lesioned skin from dogs with stage I disease was more frequently characterised by the nodular to diffuse pattern and granuloma formation (P = 0.0166) and by a lower parasite load studied by means of qPCR (P = 0.043) compared with more diseased dogs., Conclusions: Normal-looking skin from dogs with stage I is less likely to present histological lesions as well as harbour the parasite when compared with dogs with moderate to severe leishmaniosis.

The enzyme adenosine deaminase (ADA) is critical for modulating the immune system, and in the presence of zinc, its activity is catalyzed. The aim of this study was to evaluate the ADA activity in pancreas of cattle naturally infected by Eurytrema coelomaticum in relation to the results of zinc levels, pathological findings and parasite load. For this study 51 slaughtered cattle were used. The animals were divided into two groups: Group A consisting of animals naturally infected by E. coelomaticum (n=33) and Group B of uninfected animals (n=18). Blood and pancreas were collected of each animal for analysis of zinc and ADA, respectively. Infected cattle showed a reduction on seric levels of zinc, and decreased ADA activity in the pancreas (P>0.05). A positive correlation between zinc levels and ADA activity was observed. Thus, high parasite load and severity of histopathologic lesions affect the ADA activity in pancreas, as well as the zinc levels in serum of infected animals (negative correlation between these variables). Therefore, we can conclude that cattle infected by E. coelomaticum have low ADA activity in pancreas, which can be directly related to zinc reduction, responsible for ADA activation and catalyzes., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Quantification of Leishmania infantum load via real-time quantitative polymerase chain reaction (qPCR) in lymph node aspirates is an accurate tool for diagnostics, surveillance and therapeutics follow-up in dogs with leishmaniasis. However, qPCR requires infrastructure and technical training that is not always available commercially or in public services. Here, we used a machine learning technique, namely Radial Basis Artificial Neural Network, to assess whether parasite load could be learned from clinical data (serological test, biochemical markers and physical signs). By comparing 18 different combinations of input clinical data, we found that parasite load can be accurately predicted using a relatively small reference set of 35 naturally infected dogs and 20 controls. In the best case scenario (use of all clinical data), predictions presented no bias or inflation and an accuracy (i.e., correlation between true and predicted values) of 0.869, corresponding to an average error of ±38.2 parasites per unit of volume. We conclude that reasonable estimates of L. infantum load from lymph node aspirates can be obtained from clinical records when qPCR services are not available., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The genetics of tick infestation in sheep need study, as host resistance often forms part of integrated pest control programs. Repeated udder health scores, site-specific tick count, mating weight and reproduction records (N=879-1204) were recorded annually from 2010 to 2015 on ewes of the indigenous Namaqua Afrikaner (NA) fat-tailed breed, as well as the commercial Dorper and SA Mutton Merino (SAMM) breeds. Udders were scored subjectively on a 1-5 scale (1 - udder intact and 5 - udder damaged severely) and ticks were counted on three locations. The body sites counted were the head and thoracic limb (HTLTC), udder-pelvic limb (UPLTC) and perineum-breech-tail (PBTTC). These counts were also totaled for a total tick count (TTC). Reproduction traits were number of lambs weaned per ewe lambed and total weight of lamb weaned per ewe lambed. Udder health scores of NA ewes were lower than those of Dorpers, which in turn had lower scores than SAMM ewes. NA ewes had lower values for HTLTC, UPLTC and TTC than the commercial breeds, but higher values for PBTTC than Dorpers. Heritability estimates amounted to 0.26±0.04 for HTLTC, 0.53±0.04 for UPLTC, 0.07±0.06 for PBTTC, 0.44±0.06 for TTC and 0.61±0.03 for udder health score. Animal permanent environment also affected PBTTC (0.14±0.07). Significant genetic correlations were found between the HTLTC and UPLTC (0.47±0.10), UPLTC and udder health score (0.52±0.07), HTLTC and UPLTC (0.24±0.11) as well as UPLTC and PBTTC (-0.44±0.11). Heavier ewes had higher UPLTC (0.38±0.09), TTC (0.33±0.09) and impaired udder health (0.21±0.08). Udder health scores and tick counts at all sites were not related to reproduction traits. The indigenous NA breed outperformed the commercial breeds with lower values for HTLTC, UPLTC, TTC and a better udder health score. Mechanisms contributing to the better performance of the NA breed under pastoral conditions and the scope for selection for tick tolerance within breeds should be studied further., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Innate immunity, in particular, the role of toll-like receptors (TLRs), has not been extensively studied in canine L. infantum infection. The main aim of this study was to determine the transcription of TLR2 and TLR4 in the blood of dogs with natural clinical leishmaniosis at the time of diagnosis and during treatment follow-up and subsequently correlate these findings with clinical, serological and parasitological data. Forty-six Leishmania-seropositive sick dogs with a high antibody level at the time of diagnosis were studied and compared with 34 healthy seronegative dogs. Twenty-two of these sick dogs were treated with meglumine antimoniate and allopurinol and followed-up at 30, 180 and 365days following the start of treatment. Clinical status was defined by a thorough physical examination, complete blood count, biochemistry profile, electrophoresis of serum proteins, and urinary protein/creatinine ratio (UPC). EDTA blood was stored in RNAlater ® solution before RNA extraction and cDNA production were performed. TLR2, TLR4 and three reference genes (HPRT-1, CG14980 and SDHA) were studied in each blood sample by real time PCR. The relative quantification of TLR2 was higher (mean 3.5) in sick dogs when compared with seronegative healthy dogs (mean 1.3; P=0.0001) while the relative quantification of TLR4 was similar in both groups. In addition, the relative quantification of TLR2 significantly decreased during follow-up at all time points compared with day 0 whereas no changes were observed with TLR4 transcription. A significant positive correlation was noted between TLR2 and UPC, total protein, beta and gamma globulins, specific L. infantum antibodies and blood parasite load while a negative correlation was observed with albumin, albumin/globulin ratio, hematocrit and hemoglobin. TLR4 transcript did not correlate with any parameter. These findings indicate an up-regulation of TLR2 transcription in unstimulated blood in naturally infected sick dogs as compared to healthy dogs suggesting active innate immune and proinflammatory responses. In addition, TLR2 transcription is reduced with clinical improvement during treatment. In contrast, TLR4 transcription appears to be similar among groups at the time of diagnosis with no changes during treatment follow-up suggesting a less important role for this TLR in clinical canine leishmaniosis., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Intensity of peripheral parasite infection has an important role in the transmission of Leishmania spp. from one host to another. As parasite load quantification is still an expensive procedure to be used routinely in epidemiological surveillance, the use of surrogate predictors may be an important asset in the identification of dogs with high transmitting ability. The present study examined whether common clinical and laboratory alterations can serve as predictors of peripheral parasitism in dogs naturally infected with Leishmania spp. Thirty-seven dogs were examined in order to establish correlations between parasite load (PL) in multiple peripheral tissues and common clinical and laboratory findings in canine visceral leishmaniasis (CVL). Quantitative polymerase chain reaction was employed to determine PL in conjunctival swabs, ear skin, peripheral blood and buffy coat. Additionally, a series of hematological, biochemical and oxidative stress markers were quantified. Correlations between net peripheral infection and severity of clinical alterations and variation in laboratory parameters were assessed through a new analytical approach, namely Compressed Parasite Load Data (CPLD), which uses dimension reduction techniques from multivariate statistics to summarize PL across tissues into a single variable. The analysis revealed that elevation in PL is positively correlated with severity of clinical sings commonly observed in CVL, such as skin lesions, ophthalmic alterations, onycogriphosis, popliteal lymphadenomegaly and low body mass. Furthermore, increase in PL was found to be followed by intensification of non-regenerative anemia, neutrophilia, eosinopenia, hepatic injury and oxidative imbalance. These results suggest that routinely used clinical and laboratory exams can be predictive of intensity of peripheral parasite infection, which has an important implication in the identification of dogs with high transmitting ability., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Predators affect prey populations not only through direct predation, but also by acting as definitive hosts for their parasites and completing parasite life cycles. Understanding the affects of parasitism on prey population dynamics requires knowing how their predators' parasite community is affected by diet and prey availability. Ungulates, such as moose (Alces americanus) and white-tailed deer (Odocoileus virginianus), are often important prey for wolves (Canis lupus), but wolves also consume a variety of alternative prey, including beaver (Castor canadensis) and snowshoe hare (Lepus americanus). The use of alternative prey, which may host different or fewer parasites than ungulates, could potentially reduce overall abundance of ungulate parasites within the ecosystem, benefiting both wolves and ungulate hosts. We examined parasites in wolf carcasses from eastern Manitoba and estimated wolf diet using stable isotope analysis. Taeniidae cestodes were present in most wolves (75%), reflecting a diet primarily comprised of ungulates, but nematodes were unexpectedly rare. Cestode abundance was negatively related to the wolf's δ(13) C value, indicating diet affects parasite abundance. Wolves that consumed a higher proportion of beaver and caribou (Rangifer tarandus), estimated using Bayesian mixing models, had lower cestode abundance, suggesting the use of these alternative prey can reduce parasite loads. Long-term consumption of beavers may lower the abundance of adult parasites in wolves, eventually lowering parasite density in the region and ultimately benefiting ungulates that serve as intermediate hosts. Thus, alternative prey can affect both predator-prey and host-parasite interactions and potentially affect food web dynamics., (© 2016 The Authors. Journal of Animal Ecology © 2016 British Ecological Society.)

The objective of the present study was to analyze the skin (nasal surface and ear regions), lymph nodes (popliteal and pre-scapular), spleen and liver of dogs with visceral leishmaniasis (VL), in order to investigate the relationship between the parasite load measured as DNA copy number of Alpha gene of DNA polymerase of Leishmania infantum by quantitative PCR and the number of M2 macrophages by immunohistochemistry. A set of 29 naturally infected dogs from an endemic area for VL were sampled and another set of six dogs negative for VL and from a non-endemic area were analyzed as the control group (C). The spleen presented the highest number of Leishmania DNA copies, with significant differences between the groups G1 and G2 (with and without skin lesions, respectively). The M2 phenotype immunostaining predominated among the macrophages in granulomas and inflammatory infiltrates of samples from the skin, lymph nodes and spleens examined. The presence of M2 macrophages in dogs from infected group differed significantly from the control group, in all organs analyzed, excepted liver. The highest proportion of M2 macrophages coincided with the highest parasitism loads found in more susceptible organs of VL dogs, even in the skin, considered a more resistant organ, while the liver showed low parasitism load and low immunostaining for M2 macrophages with no significant differences between infected and negative groups. It was concluded that the predominance of M2 phenotype in VL dogs favored the multiplication of Leishmania infantum in organs of dogs that are more susceptible to Leishmania infection, as skin, lymph nodes and spleen., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Canine leishmaniasis treatment focuses on the reduction of parasite load, the clinical improvement of the animal, and the avoidance of relapses, in a scenario where the definitive parasite clearance is not achievable. Therefore, monitoring is crucial during the treatment of this disease. Quantitative PCR has been shown as an ideal tool for the treatment monitoring when quantifying parasite load in target organs such as lymph node or bone marrow, tissues that are too invasive for regular evaluation. This study aims to prove the potential of hair parasite load in the treatment monitoring of canine leishmaniasis. Six dogs were treated with meglumine antimoniate and monitored up to four months after the end of the treatment. Parasite loads in bone marrow, blood, lymph node and hair were quantified by real-time quantitative PCR. Total IgG, IgG1, and IgG2 antibody titres were analysed by immunofluorescent assay and a clinical assessment was carried out. Treatment consisted of two 28-day courses of meglumine antimoniate (100mg/kg/day) separated by an one-month interval. Analyses were performed before (day 0), during (day 60) and after treatment (day 120), and at the end of a follow-up period (day 210, four months after the end of treatment). Hair parasite load turned out to be strongly correlated with bone marrow, lymph node and blood parasite loads and with the clinical score and the IgG1 antibody titre. The evolution of this biomarker reflects the evolution of the parasitological, immunological and clinical state of the dog, highlighting its potential as a non-invasive marker for the treatment monitoring in canine leishmaniasis., (Copyright © 2016 Elsevier B.V. All rights reserved.)