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Lameness in pigs is a major welfare and economic issue for swine breeding herds. Claw lesions have been suggested to be a significant cause for lameness in sows. Housing conditions and nutrition management on the farm influence horn quality and may be associated with the development of claw lesions in pigs. The current work examines the structure as well as hardness, fracture and mechanical properties of claws retrieved from housing sows. For the mechanical characterisation of pig claws, an experimental program that includes three point bending test in claws was performed in order to obtain the resistance of the tissue under bending forces. The study also includes hardness measurements through Vickers method as well as inspection of the structure through Scanning Electron Microscopy. Experimental measurements show that the claw specimen behaves as a linear viscoelastic material. Measurements of hardness were found to be affected by the moisture content of the claws.

The optimal microscopic magnification and number of optical fields of adhesive tape strip cytological slides that should be examined when searching for Malassezia yeasts on canine skin are unknown.To determine the optimal magnification and the minimum number of optical fields that should be examined to maximise intraobserver repeatability and interobserver reproducibility.Seven experienced examiners counted, twice, the number of yeasts in 10, 20, 30, 40 and 50 optical fields of 40 slides at ×400 and ×1000 magnification.The number of yeasts per unit surface area was significantly higher at ×1000 compared to ×400 magnification. Repeatability and reproducibility for counting the yeasts was very poor.Adhesive tape strip cytological slides should be examined microscopically for Malassezia spp. at ×1000 magnification. The repeatability of this examination for counting the yeasts is poor.Contexte - Le grossissement microscopique optimal et le nombre de champs optiques des lames cytologiques de bandes adhésives à examiner lors de la recherche de levures Malassezia sur la peau de chien sont inconnus. Objectifs - Déterminer le grossissement optimal et le nombre minimal de champs à examiner pour maximiser la répétabilité intra-observateur et la reproductibilité inter-observateur. Matériels et méthodes - Sept examinateurs expérimentés ont compté, deux fois, le nombre de levures dans 10, 20, 30, 40 et 50 champs de 40 lames aux grossissements ×400 et ×1 000. Résultats - Le nombre de levures par unité de surface était significativement plus élevé au grossissement ×1 000 par rapport au grossissement ×400. La répétabilité et la reproductibilité du comptage des levures étaient très médiocres. Conclusions et pertinence clinique - Les lames cytologiques de bandes adhésives doivent être examinées au microscope pour Malassezia spp. à un grossissement ×1 000. La répétabilité de cet examen de comptage des levures est faible.Introducción- se desconoce el aumento microscópico óptimo y el número de campos ópticos de los portaobjetos citológicos en tiras de cinta adhesiva que deben examinarse al buscar levaduras Malassezia en la piel canina. Objetivos- determinar el aumento óptimo y el número mínimo de campos ópticos que deben examinarse para maximizar la repetibilidad intraobservador y la reproducibilidad interobservador. Materiales y métodos- siete examinadores experimentados contaron dos veces el número de levaduras en campos ópticos de 10, 20, 30, 40 y 50 de 40 portaobjetos con aumentos de x ×400 y ×1000. Resultados- el número de levaduras por unidad de superficie fue significativamente mayor con un aumento de ×1000 en comparación con un aumento de ×400. La repetibilidad y reproducibilidad para contar las levaduras fue muy pobre. Conclusiones y relevancia clínica - Los portaobjetos citológicos en tiras de cinta adhesiva deben examinarse microscópicamente para detectar Malassezia spp. con un aumento de ×1.000. La repetibilidad de este examen para contar las levaduras es pobre.Hintergrund - Die optimale mikroskopische Vergrößerung und die Anzahl der optischen Felder von zytologischen Präparaten mit Klebestreifen, die auf der Suche nach Malassezia Hefen auf Hundehaut untersucht werden sollten, sind nicht bekannt. Ziele - Eine Feststellung der optimalen Vergrößerung und der Mindestanzahl der optischen Felder, die untersucht werden sollten, um die Intra-Beobachter Reproduzierbarkeit und die Inter-Beobachter Reproduzierbarkeit maximal zu verbessern. Material und Methoden - Sieben geübte UntersucherInnen zählten zweimal die Anzahl der Hefepilze in 10, 20, 30, 40 und 50 optischen Feldern auf 40 Objektträgern bei einer Vergrößerung von x400 und x1000. Ergebnisse - Die Anzahl der Hefen pro Einheitsoberfläche war bei der x1000 Vergrößerung signifikant höher als bei der x400 Vergrößerung. Die Reproduzierbarkeit und Vergleichbarkeit der Zählungen der Hefen war sehr gering. Schlussfolgerungen und klinische Bedeutung - Zytologische Klebestreifen Präparate sollen mikroskopisch auf Malassezia spp. bei einer x1000 Vergrößerung untersucht werden. Die Wiederholbarkeit dieser Untersuchung ist in Bezug auf die Anzahl der Hefepilze schlecht.背景- 犬の皮膚に存在するマラセチア酵母を検索する際に、粘着テープ細胞診スライドの最適な顕微鏡倍率と検査すべき光学野の数は不明である。 目的-本研究の目的は、観察者内再現性および観察者間再現性を最大にするために、最適な顕微鏡倍率と観察すべき最小視野数を決定することであった。 材料と方法 - 経験豊富な7名の検査者が、40枚のスライドをx400およびx1,000の倍率で10、20、30、40、50視野の酵母数を2回カウントした。 結果-単位表面積当たりの酵母数は、400倍と比較して1,000倍で有意に多かった。酵母を数えるための反復性、再現性は非常に悪かった。 結論・臨床的意義-粘着テープ細胞診スライドは、1,000倍でマラセチア属菌の顕微鏡検査を行う必要がある。本検査では、酵母の計数の再現性が低かった。.背景-在犬皮肤上寻找马拉色菌酵母菌时, 应检查胶带条细胞学载玻片的最佳显微镜放大倍数和光野数量尚不清楚。 目的-确定最佳放大倍率和应检查的最小光野数量, 以使观察者内重复性和观察者间重现性最大化。 材料和方法-7名经验丰富的检查者在x×400和x 1,000放大倍数下, 对40个载玻片的10、20、30、40和50个光学视野中的酵母菌数量进行了两次计数。 结果-与放大400倍相比, 放大1000倍时单位表面积的酵母菌数量显著更高。计数酵母菌的重复性和重现性极差。 结论和临床相关性-应在显微镜下检查胶带条细胞学载玻片的马拉色菌。放大1000倍。该检查计数酵母菌的重复性较差。.Contexto - A ampliação microscópica ideal e o número de campos ópticos das lâminas citológicas de fita adesiva que devem ser examinados nas pesquisas de leveduras do gênero Malassezia em cães são desconhecidos. Objetivos - Determinar a magnificação ideal e o número mínimo de campos ópticos que devem ser examinados para maximizar a repetibilidade intraobservador e a reproducibilidade interobservador. Materiais e métodos - Sete examinadores experientes contaram duas vezes o número de leveduras em 10, 20, 30, 40 e 50 campos ópticos de 40 lâminas nas magnificações de x400 e x1000. Resultados - O número de leveduras por unidade de área de superfície foi significativamente maior em x1000 em comparação com a ampliação de x400. A repetibilidade e a reprodutibilidade para a contagem de leveduras foi muito pobre. Conclusões e relevância clínica - Lâminas de citologia por fica adesiva devem ser examinadas microscopicamente para Malassezia spp a uma magnificação de x1.000. A repetibilidade deste exame para contagem de leveduras foi pobre.

Feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) are retroviruses causing significant morbidity and mortality in cats. The aim of this study was to describe the epidemiological, clinical and clinicopathologic aspects of FeLV and FIV infections in different populations of cats in Greece, including client-owned cats, stray cats and cats who live in catteries. A total of 435 cats were prospectively enrolled. Serological detection of FeLV antigen and FIV antibody was performed using a commercial in-house ELISA test kit. The results showed that 17 (3.9 %) and 40 (9.2 %) of the 435 cats were positive for FeLV antigen and FIV antibody, respectively, whereas 5 (1.1 %) had concurrent infection with FeLV and FIV. Factors that were associated with FeLV antigenemia, based on multivariate analysis, included vomiting, rhinitis, infection with FIV, neutropenia, decreased blood urea nitrogen and increased serum cholesterol and triglyceride concentrations. Factors associated with FIV seropositivity included male gender, older age, outdoor access, weight loss, fever, gingivostomatitis, skin lesions and/or pruritus and hyperglobulinemia. Various clinical signs and laboratory abnormalities were found to be significantly associated with retroviral infections, suggesting that current guidelines to test all sick cats should be followed, taking into particular consideration the high-risk groups of cats found in this study.

Background Urinary tract infections (UTI) of sows characterized by cystitis, which may progress to ureteritis and pyelonephritis, can affect their productivity, longevity and welfare. In this study, we determined the prevalence of UTI by histopathology and bacteriology. Furthermore, we investigated possible associations between histologically confirmed cystitis and the results of urinalysis and urine cultures in culled sows from three farrow-to-finish herds in Greece. Materials and methods One hundred eighty-five routinely culled sows were included in the study. Their urinary bladder was collected from abattoirs and subjected to histopathology. Furthermore, urinalysis and urine cultures were performed on urine samples aseptically collected from the bladders. Results Histologically confirmed cystitis was evident in 85/185 (45.94%) culled sows. Among those, 44 (51.76%) suffered from acute and 41 (48.24%) from chronic inflammation. The majority of positive urine cultures were due to colonization of the urinary tract with E.coli, which accounted for 55.81% of the total cases, followed by Staphylococcus spp. which accounted for 18.60% of detected infections. Evidence of cystitis was associated with bacteriuria and sows with bacteriuria were 2.30 (P = 0.03, 95% CI: 1.10–4.83) times more likely to have histologically confirmed cystitis compared to sows with negative urine cultures. Bacteriuria was associated with proteinuria (P P P P P P P P = 0.006, 1.39–7.69) times less likely for a sow to have chronic or acute cystitis compared to absence of cystitis. Conclusion In the studied population, UTI affected almost one out of two culled sows. Bacteriuria, which was more common among sows with UTI than those without, was mainly ascribed to members of the intestinal and environmental bacteria. Proteinuria and the existence of urine sediment which were associated with UTI, could be used as proxy traits for UTI in live sows.

African swine fever (ASF) poses a major threat to swine health and welfare worldwide. After several European countries have reported cases of ASF, Greece confirmed officially the first positive case on 5 February 2020. The owner of a backyard farm in Nikoklia, a village in Serres regional unit, Central Macedonia, reported a loss of appetite, weakness, dyspnea, and the sudden death of 6 domestic pigs. Necropsy was performed in one gilt and findings were compatible with acute to subacute septicemic disease. Predominantly, hyperemic enlargement of spleen and lymph node enlargement and/or hemorrhage were observed. Description of vague clinical signs by the farmer suggested a limited resemblance to ASF-acute infection. However, the disease could not be ruled out once septicemic condition including splenomegaly, was diagnosed macroscopically at necropsy. In addition, considering the farm’s location near to ASF protection zones, a further diagnostic investigation followed. Confirmation of the disease was obtained using a series of diagnostic tests on several tissue samples. Further clinical, molecular, and epidemiologic evaluation of the farm was performed. According to the contingency plan, authorities euthanized all 31 pigs on the farm, whilst blood testing revealed ASF virus infection. Further emergency measures were implemented to contain the spread of the disease.

Paratuberculosis is a disease affecting mainly ruminants, and it is caused by Mycobacterium avium subspecies paratuberculosis (MAP). Control of the disease relies primarily on test-and-removal that is not easy to apply effectively for reasons associated with its pathogenesis. Therefore the aim of this preliminary study was to investigate whether positivity of sheep to MAP-specific antibody and DNA, detected by ELISA and faecal real time polymerase chain reaction (PCR) respectively, varies between specific stages of breeding i.e. before and after parturition, and before the mating season. Rectal faeces (n = 120) and whole blood (n = 120) samples were collected from a flock of sheep with no record of clinical paratuberculosis. The sheep under study are maintained throughout the year, in isolated confinement with no access to pasture or other animals, and a continuously monitored diet. The proportion of positive animals to real time PCR before parturition (13.9%) was found to be significantly lower (χ 2 = 10.67, P = 0.0015 ) than the proportion of positive animals after parturition (59.5%), and before the mating season (47.6%). The proportion of positive animals after parturition and before the mating season did not differ significantly. PCR analysis of one sample collected from each animal after parturition, allowed detection of more than 71% of the reactors. In conclusion, faecal shedding of MAP detected by real time PCR seems to be decreased before parturition, which probably makes the specific period less suitable for detection of shedders in sheep with no clinical signs of paratuberculosis.

Canine leishmaniosis (CanL)-associated chronic kidney disease is a leading cause of morbidity and mortality in Mediterranean countries. Novel renal biomarkers, such as serum symmetric dimethylarginine (sSDMA), may be useful surrogates for the detection of renal functional impairment. The objectives of this study were to investigate sSDMA concentrations in dogs with CanL, with and without azotemia, and to establish any potential association with the prevalence and severity of proteinuria, with the prevalence of decreased urine specific gravity and with the LeishVet clinical stages of CanL. Serum samples from 68 dogs with CanL (50 nonazotemic and 18 azotemic) and 17 healthy dogs were retrospectively examined. Increased sSDMA was documented in 26 % of dogs with CanL without azotemia and in 83.3 % of dogs with azotemia. Serum SDMA was significantly higher in azotemic compared to nonazotemic dogs and was associated with the presence and severity of proteinuria, the decreased urine specific gravity and the advanced clinical stages of CanL. The results of the present study indicate that sSDMA may be a useful adjunct to serum creatinine and urine protein/creatinine ratio for the detection of CanL-associated nephropathy, but it is of limited value for distinguishing among the LeishVet clinical stages of CanL.

Canine leishmaniosis due to Leishmania infantum is a widespread zoonotic disease. Although aminosidine can be an effective treatment, current therapeutic recommendations do not advocate its use, mainly due to concerns regarding the potential nephrotoxicity and ototoxicity of this drug. The aim of this randomized, blinded, controlled study was to evaluate the nephrotoxicity and ototoxicity of aminosidine-allopurinol combination and compare it with that of meglumine antimonate-allopurinol combination in non-azotemic dogs with leishmaniosis. Forty dogs with leishmaniosis were randomly assigned to be treated with either aminosidine at 15 mg/kg, subcutaneously, once daily for 28 days (group A) or with meglumine antimonate at 100 mg/kg, subcutaneously, once daily for 28 days (group B). In addition to either drug, dogs in both groups were administered allopurinol at 10 mg/kg per os twice daily for 2 months. Kidney function was evaluated through measurement of serum creatinine, urea nitrogen, inorganic phosphorus, and cystatin-c concentrations and complete urinalysis, including protein-to-creatinine ratio, at baseline and after 14, 28, and 60 days from the beginning of the treatment. At the same time points, vestibular and auditory functions were evaluated through neurological examination and brainstem auditory evoked response (BAER) recordings of wave I, wave V, inter-wave I–V latencies, and minimum hearing thresholds. None of the dogs developed clinicopathological evidence of kidney disease during the study. Serum creatinine concentration increased >0.3 mg/dl over baseline in 2 dogs in group A and in 5 dogs in group B. Parameters of kidney function were not significantly different or were improved compared to baseline and the only difference between the two groups was the lower concentration of serum creatinine in group A. None of the dogs developed peripheral vestibular syndrome or hearing impairment. At the end of the study, parameters of auditory function were not significantly different or were improved compared to baseline and there were no differences between the two groups. The results of this study show that the nephrotoxicity and ototoxicity of aminosidine, when administered to non-azotemic dogs with leishmaniosis at 15 mg/kg subcutaneously once daily for 28 days along with allopurinol, is minimal and does not differ from that of meglumine antimonate.

In this cross-sectional study we identified flock-level risk factors for Mycobacterium avium subsp. paratuberculosis (MAP) infection, in Greek dairy goat flocks. We collected 1599 milk samples from does that were at the last stage of lactation in 58 randomly selected dairy goat flocks, during May to September 2012. The collected samples were tested with a commercial milk ELISA (IdexxPourquier, Montpellier, France) and the results were interpreted at a cut-off that optimized the accuracy of the diagnostic process. For the analysis of the data we used Bayesian models that adjusted for the imperfect Se and Sp of the milk-ELISA. Flock was included as a random effect. Does in flocks that used common water troughs and communal grazing grounds had 4.6 [95% credible interval (CI): 1.5; 17.4] times higher odds of being MAP-infected compared to does in flocks that had no contact with other flocks. Does of flocks supplied with surface water from either streams or shallow wells had 3.7 (1.4; 10.4) times higher odds of being infected compared to those in flocks watered by underground and piped water sources. When kids were spending equal to or more than 10h per day with their dams they had 2.6 (1.1; 6.4) times higher odds of being MAP infected compared to kids that were separated from their dams for less than 10h per day. Finally, does in flocks that continuously used the same anti-parasitic compound had 2.2 (1.0; 4.6) times higher odds of MAP infection compared to those in flocks alternating anti-parasitic compounds. These results should be considered in the development of a nationwide future control program fοr caprine paratuberculosis in Greece.

Natural infection of domestic cats by Leishmania infantum (synonym: L. chagasi) has been demonstrated in several European, Latin American, and Asian countries, and the estimated prevalence of infection, based mainly on blood PCR, ranges from 0.3% up to 60.6%. In this study we aimed to: (a) estimate the prevalence of the infection by L. infantum in clinically normal cats (group A; n = 50) and in cats with various clinical signs (group B; n = 50), living in an endemic region, by both cytological examination of four different tissues (lymph node, skin, bone marrow, and conjunctiva) and by PCR in four different tissues (blood, skin biopsies, bone marrow, and conjunctiva); (b) compare the diagnostic sensitivity of the above methods and evaluate for possible associations between their results; and (c) investigate the possible associations between infection by L. infantum and signalment, living conditions, season of sampling, and health status of the cats. The prevalence of the infection in the study population was 41% and did not differ (P = 0.839) between group A (42%) and B (40%) cats. Lymph node, skin, bone marrow and conjunctiva cytology was always negative. Therefore, the diagnosis of the infection was based only on PCR in blood, skin biopsy, bone marrow and conjunctiva, which was positive in 13%, 18.2%, 16% and 3.1% of the cats, respectively. PCR was positive in only one of the four tissues in 80.5% of the infected cats. The results differed (P = 0.014) among the four tissues and were less frequently positive in conjunctiva compared to skin biopsies and bone marrow (P = 0.007 for both comparisons), thus highlighting the need for multiple tissue PCR testing in order to minimize false-negative results. More PCR-positive cats were found when sampling was performed during the period of sandfly activity (odds ratio: 2.44; P = 0.022). Also, in group B cats, the likelihood of PCR-positivity was higher (odds ratio: 3.93; P = 0.042) among those presenting at least one systemic clinical sign that had been previously reported in cats with leishmaniosis.

Background Hoof lesions in sows have been associated with lameness and poor hoof horn quality. The mechanical strength and quality of hoof horn is determined by the density and diameter of horn tubules, which were recently associated with the severity of lesions on the hoof wall of sows. Histologic changes that have previously been described in cases of bovine laminitis, have also been observed in the dermis and epidermis of the sows’ claws. Trace elements, particularly zinc, copper and manganese, occupy important roles as enzyme catalysts in the process of keratin synthesis which determines the quality and the integrity of the hoof epidermis. Therefore, the objective of this study was to investigate the effect of diet supplementation with chelated zinc, copper and manganese, partially substituting their inorganic form, on sow claw health and hoof horn quality assessed by macroscopic, histologic and morphometric examination. Results Clinically, the total claw lesion score was significantly lower in claws of sows which received the “organic” diet compared to those of sows on the “inorganic” diet. Histologically, lamellar hyperplasia was the most frequently recorded change in the epidermis of the sows’ claws regardless of the diet’s mineral source. The claws of the sows which received the organic diet were more likely to have none or less histologic changes than at least one or more, respectively, compared to those of the sows on the “inorganic” diet. Morphometrically, the density and vertical and horizontal diameters of the horn tubules was significantly higher and smaller, respectively, in the hoof horn of sows which received the “organic” compared to those which received the “inorganic” source diet. Conclusions Partial substitution of the inorganic zinc, copper and manganese in sows’ diet with their chelated complexes, provided a comparative advantage against a conventional, inorganic mineral source diet, at least under the conditions examined in the current study, in terms of macroscopic, histologic and morphometric criteria, characterizing the health and horn quality status of sows’ hooves.

Canine spirocercosis (CS) is a helminthic infection caused by the nematode Spirocerca lupi. The clinical hallmark of the disease is esophageal dysphagia due to parasite-induced esophageal nodules. Currently, there is limited information on the involvement of serum acute phase proteins (APPs) in the symptomatic CS. The objective of this study was to investigate whether C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp) and albumin are involved in CS, and if their concentrations measured on admission reflect the severity of benign esophageal lesions. Nineteen dogs with spontaneous symptomatic esophageal spirocercosis and 7 clinically healthy dogs were studied retrospectively. The most consistently increased APP in the symptomatic dogs was Hp (95% of the dogs), followed by CRP (68%). The SAA concentrations were infrequently increased (5% of the dogs), while albumin concentrations were decreased in 58% of the affected dogs. The dogs with spirocercosis had significantly higher median concentrations of Hp (p=0.0001) and CRP (p=0.02) compared to healthy dogs. Median albumin concentrations did not differ between the two groups of dogs. The median concentrations of Hp, CRP and albumin did not differ significantly between the dogs having a single or multiple esophageal nodules. The results of this study indicate that in symptomatic CS, Hp and CRP are significantly and consistently increased, while SAA and albumin may be of limited value as diagnostic markers. No association was established between the concentrations of Hp, CRP and albumin measured on admission and the number of esophageal nodules.

Background: Canine monocytic ehrlichiosis (CME), caused by Ehrlichia canis, is an important tick-borne disease of global importance. Currently, limited information is available on the diagnostic and prognostic value of acute phase proteins (APPs) in dogs naturally infected with E. canis. Hypothesis: APPs may be useful indicators of the clinical phase of CME and predictive of the clinical outcome (death or survival). Animals: Fifty-six dogs naturally infected with E. canis and 7 clinically healthy control dogs. Methods: C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp), and albumin concentrations determined on admission were retrospectively compared among 27 dogs with nonmyelosuppressive CME, 29 dogs with myelosuppressive CME and 7 healthy dogs. Diagnosis of CME was based on clinical and clinicopathological findings, seropositivity to E. canis, polymerase chain reaction amplification of E. canis-specific 16S rDNA, microscopic observation of Ehrlichia sp. morulae in blood monocytes or some combination of these. Results: Mean concentrations of CRP, SAA, and Hp were significantly higher in the myelosuppressed dogs compared with the other groups, but no significant differences were found in the concentration of albumin. Survival analysis of the affected animals indicated that APP concentrations were not associated with clinical outcome; the latter was strongly associated with pancytopenia (odds ratio for death 22.7) and neutropenia (odds ratio for death 7.7). Conclusions and Clinical Importance: CRP, SAA, and Hp serum concentrations on admission are useful indicators of the clinical phase of CME, but are not useful predictors of clinical outcome.

Background: Recognition of different cytologic patterns in lymph nodes (LNs) from dogs with canine monocytic ehrlichiosis (CME) and noninfectious causes of lymphoid reactivity may have diagnostic utility. Objectives: The aims of the present study were to compare cytologic patterns in LNs of dogs with different phases of CME, to investigate the association of cytologic pattern and presence of Ehrlichia spp. morulae, and to compare patterns of lymphoid reactivity between dogs with CME and those with noninfectious causes of lymphoid hyperplasia. Methods: Cytologic preparations of LNs from 35 dogs with nonmyelosuppressive CME (group A), 16 dogs with myelosuppressive CME (group B), 26 dogs with noninfectious diseases (group C), and 15 healthy dogs (group D) were evaluated. Percentages of lymphocyte types, plasma cells, macrophages, neutrophils, and eosinophils were determined. Samples from dogs in groups A and B were evaluated for the presence of morulae. Results: Cytologic abnormalities in LNs were recorded in 54% of dogs in group A, 88% in group B, 39% in group C, and 0% in group D and were more frequent (P=.02) in dogs with myelosuppressive CME than those with nonmyelosuppressive CME. Plasma cell hyperplasia was more frequent in CME than in noninfectious diseases (P=.03). An association between the presence of cytologic abnormalities and morulae in group A dogs was not found. Conclusions: Dogs with myelosuppressive CME have more lymphoid cytologic abnormalities than dogs with nonmyelosuppressive CME. LN plasmacytosis is the major pattern of lymphadenopathy in dogs with CME and is found more frequently in dogs with CME than in dogs with noninfectious causes of lymphadenopathy.

We estimated the prevalence of Mycobacterium avium subsp. paratuberculosis (MAP) infected sheep and goats, older than 2 years, reared in the region effectively controlled by the Government of the Republic of Cyprus, in September 2005. The sera collected from 8011 animals (3429 sheep and 4582 goats) from 83 flocks, non-vaccinated against MAP, were examined for antibodies with a commercially available enzyme linked immunosorbent assay (Pourquier® paratuberculosis antibody screening ELISA). The true prevalence of MAP infection was calculated, in a Bayesian framework, separately in sheep and goats, with models that adjusted for the misclassification of animals because of the imperfect accuracy of the ELISA. The within-flock mean seroprevalence in sheep and goats was 9.9% (95% CI: 8.9; 10.9%) and 7.9% (7.2; 8.7%), respectively. There was at least one seropositive sheep in 52% (38; 66%) of flocks with sheep and one seropositive goat in 50% (39; 62%) of flocks with goats. Within the seropositive flocks the mean seroprevalence was 12.1% (10.9; 13.4%) and 10.3% (9.3; 11.4%) in sheep and goats, respectively. The calculated mean true prevalence of infected sheep and goats was 15.0 and 11.1%, respectively. There was at least one infected sheep in 60.8% (95% credible interval: 42.3; 78.8%) and at least an infected goat in 48.6% (30.4; 68.5%) of the flocks. In the infected flocks, the mean within flock true prevalence of infection in sheep and goats was estimated at 24.6% (16.3; 33.3%) and 23.1% (15.5; 33.6%), respectively. © 2010 Elsevier B.V.

Despite its extensive use for evaluation of spermatogenesis and assisted reproduction, the safety and consequences of fine (FNA) and large needle aspiration (LNA) to the testicular parenchyma and its normal function have not been established. This study was performed in order to accurately assess, by serial in vitro ultrasonographic, bacteriologic, gross anatomic and histological examinations, the type and extent of the effect of FNA or LNA on the dog's testis. Twenty three sexually mature, 1 to 2 years old, healthy laboratory Beagles were randomly assigned to 2 groups: (1) 5 dogs without testicular aspiration (control group) and (2) 18 dogs in which one of their testes was aspirated using a 23 G butterfly needle and the other using a 19 G butterfly needle (experimental group). Two dogs at a time were castrated 10 minutes, 60 minutes, 2, 14, 29, 63, 76, 90 or 180 days post-aspiration. The control group was also castrated 2, 29, 63, 90 or 180 days after the beginning of the experiment. Following castration, in vitro ultrasonographic, gross anatomic, cytological examinations of epididymal sperm, bacteriologic and histological examinations of the testes were performed. Following testicular FNA and LNA bacteriologic, gross anatomic, histologic, epididymal sperm findings and the in vitro ultrasonographic appearance of the testis were normal, except of intratesticular haemorrhage, detected the first days post-aspiration, and degeneration of less than 1.5% of the seminiferous tubules. Within the parameters of this experiment, testicular FNA and LNA have no ill effect on the canine testis and therefore, both FNA and LNA should be considered safe.

The safety and consequences of fine- (FNA) and large-needle aspiration (LNA) to the testicular parenchyma and its normal function have not been thoroughly established. This study was performed to accurately assess, by serial clinical, in vivo ultrasonographic and seminological examinations, the type and extent of the effect of FNA or LNA on canine testes. Eighteen sexually mature, 1-2 years old, healthy laboratory Beagles were used. One of their testes was aspirated using a 23-G butterfly needle (FN) and the other using a 19-G butterfly needle (LN). Two dogs at a time were orchiectomized 10, 60 min, 2, 14, 29, 63, 76, 90 or 180 days post-aspiration. Five and 2 days and 1 h before aspiration (in all dogs), immediately post-aspiration, and 1, 2, 4, 7, 9, 14, 19, 29, 35, 43, 49, 56, 63, 70, 76, 90, 111, 132 and 180 days post-aspiration (in the remaining intact dogs), evaluation of scrotal surface temperature over each testis, evaluation of scrotum-testis volume by electronic sliding callipers, ultrasonographic evaluation of testicular volume and texture and clinical and semen examination were performed. Following FNA and LNA, the clinical and ultrasonographic appearance of the testis were normal. Sperm production nearly always remained unchanged, with the exception of a slight decrease in spermatozoal motility 2-14 days post-aspiration. However, even then, with the exception of six samples, spermatozoal motility was above normal values. Within the parameters of this experiment, testicular FNA and LNA have no ill effect on sperm production or clinical and ultrasonographic appearance of the canine testis, and therefore, both FNA and LNA should be considered safe.

The objectives of this matched case-control study in a veterinary teaching hospital were to investigate the influence of signalment and historical data on the odds of occurrence of canine parvovirus (CPV) enteritis and the potential usefulness of the clinical signs and clinicopathologic abnormalities recorded on admission as prognostic indicators of mean duration of hospitalization (DOH) and outcome of the disease. Ninety-four puppies with natural CPV enteritis and 188 age-matched controls were studied. The odds to develop CPV enteritis were higher in purebreds compared to mixed-breed puppies. Vomiting and depression at the time of admission were associated with a prolongation of DOH by 2 and 1.75 days, respectively. The lymphopenic and hypoalbuminemic dogs were hospitalized for 1.9 and 2.5 more days, respectively, compared to those without these abnormalities. The odds of non-survival were higher in those puppies with evidence of systemic inflammatory response syndrome (SIRS) at the time of admission.

Survival analysis methods can be used to identify factors associated with the time to induction of infection. In the absence of a perfect test, detection of infection is generally delayed and depends on the duration of the latent infection period. We assess, via simulations, the impact of ignoring the delayed detection of infection on estimated survival times and propose a Bayesian Weibull regression model, which adjusts for the delayed detection of infection. The presence of non-differential detection delay seriously biased the baseline hazard and the shape of the hazard function. For differential detection delay, the associated regression coefficients were also biased. The extent of bias largely depended on the longevity of the delay. In all considered simulation scenarios our model led to corrected estimates. We utilized the proposed model in order to assess the age at natural infection with Mycobacterium avium subsp. paratuberculosis (MAP) in Danish dairy cattle from the analysis of available time to milk-seropositivity data that detected infection with delay. The proposed model captured the inverse relationship between the incidence rate of infection and that of seroconversion with time: susceptibility to infection decreases with time (shape parameter under the proposed model was rho=0.561), while older animals had a higher probability of sero-converting (rho=2.671, under standard Weibull regression). Cows infected earlier in their lives were more likely to subsequently shed detectable levels of MAP and, hence, be a liability to herd-mates. Our approach can be particularly useful in the case of chronic infections with a long latent infection period, which, if ignored, severely affects survival estimates.

The aim of this study was to estimate the relative diagnostic sensitivity and specificity of a polymerase chain reaction (PCR) assay in the serum of dogs with naturally occurring non-myelosuppressive canine monocytic ehrlichiosis (CME), and to investigate the association between PCR positivity and immunofluorescence antibody (IFA) titres for Ehrlichia canis. Serum samples obtained from 38 dogs with non-myelosuppressive CME and 12 healthy dogs were analyzed retrospectively. Each serum sample was analyzed in triplicate using an E. canis-specific nested PCR assay targeting a 389 bp sequence of the 16S rRNA gene. E. canis DNA was amplified in 24 of 38 (63.1%) affected dogs; all samples from healthy dogs were negative. A high level of agreement was found among the PCR replicates (P < 0.0001). Median IFA titre of the 24 PCR-positive dogs was significantly lower than that of the PCR-negative infected dogs (P = 0.0029), indicating that E. canis DNA may circulate prior to the development of a high antibody titre. Serum-based PCR analysis is suggested for the early diagnosis of CME when whole blood samples are not available.

The variance partition coefficient (VPC) measures the clustering of infection/disease among individuals with a specific covariate pattern. Covariate-pattern-specific VPCs provide insight to the groups of individuals that exhibit great heterogeneity and should be targeted for intervention. VPCs should be taken into consideration when planning study designs, modeling data and estimating sample sizes. We present a Bayesian discrete mixed model for the estimation of covariate-pattern-specific VPCs when measurement of the infection/disease is based on an imperfect test. The utility of the presented model is demonstrated with three applications. In all cases, imperfect tests biased VPC estimates towards the null but corrected estimates could be obtained by modeling the sensitivity and specificity of the test procedure with beta distributions. The comparison of adjusted VPCs between the intercept only and the fitted models with higher level covariates explained the portion of heterogeneity in the data that was accounted for by the covariates.

In 2002, postweaning multisystemic wasting syndrome (PMWS) was diagnosed in a European female wild boar (Sus scrofa), based on the detection of porcine circovirus 2 (PCV2) DNA in various organs, including the uterus, and on histopathologic lesions. This is the first detection of PCV2 DNA in the uterus of a wild boar. Three years later (2005), a wild boar6-8 mo of age was found moribund. It presented wasting and dyspnea and finally died. PCV2 DNA was detected in tissue samples, and histopathologic lesions consistent with PMWS were observed. Both wild boars were from neighboring hunting areas in central Greece. Two PCV2 strains from the wild boars were genetically characterized and compared to other reported PCV2 sequences from wild boars and domestic pigs. The PCV-2 sequences from the wild boars in this study were closely related to each other and were grouped with two isolates from wild boars from Hungary. The phylogenetic analysis revealed that the virus might be transmitted between hunting areas. In addition, PCV2 may spread from domestic pigs to wild boars and vice versa.

Ο προσδιορισμός της στάθμης της κορτιζόλης eίναι η φυσιολογική παράμeτρος που χρησιμοποιeίται συχνότeρα στην eκτίμηση της eυζωίας των χοίρων. Για την αξιολόγηση της καταπόνησης (stress) των χοίρων που σταβλίζονται ομαδικά, η συλλογή δeιγμάτων ούρων κατά την ούρηση των ζώων eίναι eύκολη και πρακτική. Σκοπός της eργασίας αυτής ήταν η eφαρμογή υγρής χρωματογραφίας σe συνδυασμό μe φασματογραφία μάζας υψηλής ανάλυσης για την πeριγραφή της καμπύλης απέκκρισης κορτιζόλης στη διάρκeια της ημέρας σe ούρα ομαδικά σταβλισμένων χοιρομητέρων. Ο προσδιορισμός της κορτιζόλης έγινe σe δeίγματα ούρων που συλλέχθηκαν κατά τη διούρηση από 30 χοιρομητέρeς τριών eλληνικών eκτροφών. Από κάθe ζώο συλλέχθηκαν eπαναλαμβανόμeνα δeίγματα την ίδια ημέρα. Βρήκαμe ότι η στάθμη της κορτιζόλης στα ούρα ήταν υψηλότeρη πριν το πρωινό τάισμα των ζώων [ο γeωμeτρικός μέσος όρος του λόγου της στάθμης της κορτιζόλης προς τη στάθμη της κρeατινίνης ήταν 2.72 (95% eμπιστοσύνης: 1,17, 6,30), 5,65 (3,15, 10,14) και 2,60 (1,50, 4,50) στις χοιρομητέρeς των eκτροφών A, B, και Γ, αντιστοίχως] και χαμηλότeρη στις 7:00 μ.μ. [0,56 (0,27, 1,18), 1,24 (0,74, 2,07), 0,88 (0,55, 1,44)]. Οι καμπύλeς ημeρήσιας απέκκρισης της κορτιζόλης στα ούρα των χοιρομητέρων διέφeραν σημαντικά μeταξύ των τριών eκτροφών.

We investigated the prediction of ovarian response using oestradiol determination, in 37 gonadotrophin stimulated Karagouniko ewe-lambs. Ovarian stimulation was induced by serial FSH administrations, and laparoscopic follicular aspiration (OPU) was conducted 12h after the last FSH injection. Oestradiol concentration was assessed in six blood samples collected prior to each FSH injection and in one sample collected prior to follicular aspiration. According to ovarian response, ewe-lambs were allotted in three groups: good, L1 (n=17); moderate, L2 (n=10); and poor, L3 (n=10). Based on the data obtained from 28 (75%) randomly selected animals, a statistical model was designed and tested on the remaining nine lambs for its ability to predict the probability of good ovarian response. From the 2nd sample, oestradiol concentration was constantly higher in group L1 in comparison with L3 lambs (all p-values for the contrasts were/=0.02), while this difference between L1 and L2 lambs was significant only in the 6th and 7th sample (both p0.005). Using as criterion the oestradiol concentration of the 6th sample, the statistical model predicted all lambs that did not belong to group L1 and three of four lambs that belonged to group L1. Our results indicate that the moderate- and poor-donors could be safely predicted on the basis of oestradiol concentration 12h prior to the scheduled follicular aspiration. Moreover, poor-donors could be identified--and rejected from further manipulations--on the basis of their inability to exhibit increased oestradiol levels in response to gonadotrophin stimulus.

The objective of this retrospective study was to investigate in 100 dogs with otitis externa (OE) the possible associations between signalment, history, clinical and laboratory findings and the various primary, secondary and perpetuating causative factors of ear canal inflammation. The age of the dogs ranged from 3 months to 14 years (median: 4.75 years) and they included 45 males and 55 females. Cocker spaniels, Jura des Alpes and Brittany spaniels were significantly overrepresented among dogs with OE when compared to the hospital canine population. In the majority of the cases, OE was chronic-recurrent (63%) or bilateral (93%). Allergic dermatitis (43/100 dogs), grass awns (12/100) and otoacariasis (7/100) were the most common primary causative factors; no primary factor could be incriminated in 32 cases and more than one was found in three dogs. Malassezia spp. (66/100 dogs), cocci (38/100) and rods (22/100) were the secondary causative factors, while ear canal stenosis (38/100) and tympanic membrane perforation-otitis media (25/100) were the most important perpetuating factors. Atopic dermatitis and adverse food reactions-associated OE was more common in females and dogs with a history of pruritic skin disease, while grass awn-induced OE occurred in cocker spaniels and acute cases. Tympanic membrane perforation was less frequent in atopic dermatitis and adverse food reactions-associated OE, but more common when otoscopic and ear canal cytological examination revealed the presence of grass awns and rods, respectively. Finally, cocci overgrowth was positively associated with ear canal stenosis.

This study was designed to compare the sensitivity of deep skin scraping, hair plucking, and exudate microscopy for the diagnosis of canine demodicosis. Sixty-seven dogs diagnosed with demodicosis were enrolled in the study. Thirty dogs had localized and 37 had generalized demodicosis. Twenty-seven of the 67 dogs had complicated (secondarily infected) and 40 had noncomplicated form. On each dog, a single lesion was randomly selected to obtain one deep skin scraping, hair plucking, and, when present (n = 13) exudate. For skin scraping and exudate microscopy, an area under a cover slip measuring 2.2 x 2.2 mm was examined, while trichography included the evaluation of 100 hair shafts. At least one parasitic element was found in 85.1% of trichograms, and 100% of exudate preparations. The number of parasitic elements was higher in skin scrapings compared to the other two methods. The diagnostic sensitivity of skin scrapings was higher than that of hair pluckings for the total number of samples (P = 0.002) and for those obtained from dogs with the localized (P = 0.004) and the noncomplicated (P = 0.002) forms of the disease. The diagnostic sensitivity of hair pluckings was higher in generalized and complicated demodicosis compared to the localized and noncomplicated variants. Based on these results, exudate microscopy may be equally sensitive to deep skin scrapings, and trichography may be of value in generalized and complicated demodicosis, although a negative result cannot rule it out.

There is currently lack of information on the changes of acute phase proteins (APP) and antioxidant markers and their clinical relevance as treatment response indicators in canine monocytic ehrlichiosis (CME). The objective of this study was to investigate the patterns of C-reactive protein (CRP), haptoglobin (Hp), ferritin and paraoxonase-1 (PON-1) during treatment of dogs with acute CME with rifampicin. Blood serum samples from ten Beagle dogs with experimental acute CME were retrospectively examined. Five dogs (Group A) were treated with rifampicin (10 mg/Kg/24 h), per os, for 3 weeks and 5 dogs (Group B) received no treatment (infected controls). Two Beagle dogs served as uninfected controls. Blood serum samples were serially examined prior to Ehrlichia canis inoculation and on post-inoculation days 14, 21, 28, 35 and 42. Significant changes of CRP, Hp, ferritin and PON-1 values were found in the majority of infected dogs. However, their concentrations did not differ between the two groups during the treatment observation period. The results of this study indicate that although several APP and PON-1 tend to significantly change in the majority of dogs with acute CME, they were of limited clinical relevance as treatment response indicators in this experimental setting.

A total of 854 paired milk and blood samples were collected from ewes of a Greek flock and used to estimate the sensitivity and specificity of a commercial ELISA for detection of Mycobacterium avium subsp. paratuberculosis (MAP) specific antibodies in each stage of lactation. We implemented Bayesian mixture models to derive the distributions of the test response for the healthy and the infected ewes. In the colostrum period, early, mid and late lactation stage the median values of the area under the curves (AUC) were 0.61 (95% credible interval: 0.50; 0.84), 0.61 (0.51;0.84), 0.65 (0.51;0.91), 0.65(0.51;0.89) for the serum ELISA and and 0.60 (0.50; 0.84), 0.61 (0.50; 0.84), 0.67(0.51; 0.91), 0.66(0.50; 0.90) for the milk ELISA, respectively. Both serum and milk ELISA had low to average overall discriminatory ability as measured by the area under the curves and comparable sensitivities and specificities at the recommended cutoffs.

Latent class models were used to estimate the sensitivity (Se) and the specificity (Sp) of a serum ELISA and a faecal culture (FC) method for the diagnosis of paratuberculosis separately, in sheep and goats. The estimates were obtained by a Bayesian method. Possible dependence of diagnostic errors was investigated by comparing models where independence was assumed to models allowing for conditional dependence given the true disease status. ROC analysis for the serum ELISA was also performed and optimized cut-off values based on the misclassification cost term were determined. No evidence of conditional dependence was found. Assuming independence, posterior medians and 95% credible intervals for the Se(ELISA), Sp(ELISA), Se(FC) and Sp(FC), were 63% (42, 93%), 95% (90, 98%), 8% (2, 17%) and 98% (95, 100%) in goats and 37% (10, 80%), 97% (93, 99%), 16% (2, 48%) and 97% (95, 99%) in sheep. AUC was calculated 0.702 for sheep and 0.847 for goats. For the serum ELISA, there is need of species- and purpose-specific cut-off selection. For instance, with 20% prevalence situation and assuming equal and five-fold cost of a false negative to a false positive test result, the optimal cut-off is 0.3 and 0.05 in sheep, respectively, while it is 0.6 and 0.1 in goats, respectively. Serum ELISA performed better in goats than in sheep. Lowering the cut-off, in relation to the one recommended by the manufacturer, improved Se(ELISA) without seriously compromising Sp(ELISA), in either species.

Background Bone marrow mastocytosis has been reported rarely in naturally occurring canine monocytic ehrlichiosis (CME). Objectives The aims of the present study were to estimate the prevalence and magnitude of bone marrow mastocytosis in a case series of dogs with natural CME and to assess the association, if any, between mastocytosis and the clinical severity of the disease. Methods Seventy-six dogs with confirmed CME (Ehrlichia canis) were included in the study. Affected dogs were allocated into group A (n = 51) without bone marrow hypoplasia and group B (n = 25) with bone marrow hypoplasia. Twenty clinically healthy Beagles not previously exposed to E canis served as controls (group C). The main inclusion criteria for group A were documentation of normocellular to hypercellular bone marrow and complete clinical cure following a 4-week treatment with doxycycline, while those for group B were bone marrow hypoplasia and lack of response to doxycycline. Bone marrow aspirate smears from all 96 dogs were Giemsa-stained and examined for the presence of mast cells, which were calculated as a percentage of 1,000 nucleated cells (NCs). The prevalence of mastocytosis was compared among the 3 groups by the Pearson's chi-square test. Results Bone marrow mastocytosis (>0.1% of NCs) was found in 5 (20%) dogs in group B (range, 0.5-2.5% of NCs; median, 1% of NCs). One dog in each of groups A and C had 0.1% mast cells in the marrow. The prevalence of bone marrow mastocytosis in dogs in group B was significantly higher (P = .004) than in groups A and C. Conclusion Bone marrow mastocytosis can be seen in a substantial number of dogs with E canis-induced myelosuppression.

In this study, we compared the frequency of isolation of Mycobacterium avium subsp. paratuberculosis (MAP) from faecal samples grown on Herrold's egg-yolk medium (HEYM) or on Lowenstein–Jensen (LJ) medium and estimated the sensitivity (Se) and specificity (Sp) of the methods separately in sub-clinically infected Greek dairy sheep and goats, using latent-class models and Bayesian estimation procedures. Faecal and blood samples were collected from 400 animals ≥1 year old in April–May 2002. The HEYM supported growth of MAP better than the LJ method and their agreement was very poor (weighted kappa = 0.062 (95% CI: −0.098, 0.222)). There was no evidence of dependence between the Ses whereas the Sps were positively correlated. Thus, a semi-dependent model that assumed independence of Ses and accounted for the dependence of Sps was adopted. Under this model, the parallel interpretation of the results of the two methods gave median estimates and 95% credible intervals (CrIs) for Se par , Sp par of 15% (CrIs: 3, 45%), 96% (92, 98%) in sheep and 16% (6, 36%) and 97% (94, 99%) in goats.

Our cross-sectional study investigated the association of sub-clinical Mycobacterium avium subsp. paratuberculosis (MAP) infection with failing to produce a live offspring the season of lambing/kidding (November 2001 to January 2002) before testing (in April-May 2002), in four dairy-sheep and/or goat flocks in Greece (369 animalsor=1.5-year-old). From each selected animal 10 ml of blood and 10 g of feces from the rectum were obtained. The harvested sera were tested for antibodies to MAP with a commercial ELISA test kit; the feces were cultured on Herrold's egg-yolk medium supplemented with mycobactin J and antibiotics. An animal was considered sub-clinically infected when found either seropositive or culture positive. The true prevalence of sub-clinically infected animals, adjusted for the sensitivity and specificity of the parallel testing, was 14% (0.1-28%) and 35.9% (9.2-62.7%) in sheep and goats, respectively. The association of fertility of sheep and goats with sub-clinical paratuberculosis was investigated in random-effects logistic models. Sub-clinically infected animals (compared to uninfected) had OR for live offspring the previous year of 5.4 for parity4, OR=0.05 for parity6, and a non-significant OR for the middle parity category.

We describe the features of an outbreak of ovine abortions caused by Erysipelothrix rhusiopathiae in a flock of sheep in Greece. Abortion occurred in 3.5% of ewes in the flock; the foetuses were well developed and had congested lungs and sero-haemorrhagic fluid in the thoracic cavity; pinpoint, necrotic-like lesions were recorded on the placental cotyledons, whilst the inter-cotyledonary areas appeared normal. The same findings were also recorded in the foetus and the foetal membranes of a pregnant ewe that died and was necropsied; on the foetal membranes of that animal, bright yellow-coloured amniotic plaques were also seen. Lameness was recorded in 3% of the ewes in the flock; oedema of the periarticular tissues and sparse synovial fluid in the infected joints were evident in a ewe that died and was necropsied. Long-standing lameness was recorded in two fattening pigs, reared in the farm. E. rhusiopathiae was isolated in pure culture and heavy growth from joints of the pigs, joints, lungs, liver, foetuses and placentae of the ewe that died, aborted foetuses and placentae from the ewes that aborted, synovial fluid from the lame ewes. The identity of the organism was confirmed by means of conventional bacteriological techniques and the API-system. Specific tests for other common causes of ovine abortion were carried out, but no evidence of involvement of other agents was found. On the basis of all the evidence available, a diagnosis of erysipelas was made. Penicillin was given to all ewes in the flock and sanitation measures were applied to control the disease.

Thirty-nine Greek dairy herds, totalling 6333 cattle, enrolled in a voluntary bovine viral diarrhoea virus (BVDV) eradication programme based on the identification and removal of persistently infected (PI) animals. The aim of this study was to estimate the prevalences of BVD antigen-positive and PI animals, and investigate the significance of the associations between the prevalence estimates and herd size. Initially, all animals were bled and examined for BVDV, using an antigen ELISA. A second sample was collected from the positive animals, after a period of at least three weeks. Animals retested positive were classified as PI. Antigen positive and PI animals were detected in all herds. The respective mean prevalences, adjusted for the test's accuracy and the herd-clustering effect, were 14% (95%CI: 11-18%) and 1.3% (0.8-1.8%), respectively. Herd size was not associated with the prevalence of antigen-positive or PI animals.

The sensitivity and specificity of lymph node and bone marrow smear microscopy for the diagnosis of Leishmania infantum-infected dogs was evaluated in 79 dogs with leishmaniasis (Group A), 52 asymptomatically infected dogs (Group B), and 44 healthy noninfected dogs (Group C). Light microscopy examination included 10 to 1,000 oil immersion fields, and the density of Leishmania amastigotes was scored by a 0 to +6 scale. Using polymerase chain reaction as the gold standard, the specificity of lymph node and bone marrow cytology was 100%, whereas sensitivity ranged from 7.8% to 92.6%, being significantly higher in Group A compared with Group B. The amastigote scores were also significantly higher in Group A compared with Group B. These results indicate that lymph node and bone marrow cytology is a highly sensitive and specific method for the diagnosis of canine patent leishmaniasis, whereas its sensitivity is relatively low in asymptomatic infections.

A total of 95 clinically healthy and seronegative for Leishmania infantum dogs, residing an area highly endemic for canine leishmaniosis (CL) and living an outdoor life-style, were split into positive and negative groups, and then were randomly assigned to receive allopurinol ( n = 51; 20 mg/kg once daily), or placebo ( n = 44) for 1 week per month, from April to November. Forty per cent (38/95) of these dogs were not reexamined and retested at the end of the trial for reasons unrelated to CL. None of the remaining 57 dogs exhibited the symptomatic form of the disease at the end of the 1-year follow-up period. Of the 15 allopurinol-treated dogs that were non-infected (negative PCR and tissue smear microscopy) at the beginning of the trial, 6 (40% P = 0.03) became PCR-positive, of which 3 became also seropositive, at the end of the observation period. In contrast, only 1 of 7 (14.3%) placebo-treated non-infected dogs became PCR positive at the same time point. Of the 19 allopurinol-treated dogs that were infected (PCR-positive) at the beginning of the trial, 18 (94.7%) remained PCR-positive and one (5.3%) seroconverted, at the end of the observation period. Of the 16 initially infected and placebo-treated dogs, 14 (87.5%) remained PCR positive, of which one (6.7%) also turned positive by tissue smear microscopy. Therefore, it is concluded that the use of allopurinol, at the daily dose of 20 mg/kg, for 1 week per month, during the period of sandfly activity, does not prevent the infection of non-infected dogs by L. infantum , and, also, does not help in the elimination of the parasite from dogs with asymptomatic infections.

The prevalence of IgG antibodies to Hepatozoon canis and the presence of gamonts in the blood and hemolymphatic tissues were studied in dogs with canine monocytic ehrlichiosis (CME) caused by Ehrlichia canis. Both pathogens are transmitted by the tick Rhipicephalus sanguineus. Forty-five out of 69 (65.2%) dogs with CME were seropositive to H. canis by an enzyme-linked immunosorbent assay (ELISA). Intra-neutrophilic gamonts of H. canis were found in 2 out of 69 dogs (2.9%) comprising 4.5% of the seropositive dogs. The present study indicated that the prevalence of antibodies to H. canis was high among dogs with CME in an area where both infections are endemic. However, previous exposure to H. canis was not found as an important contributor to clinical or clinicopathologic abnormalities found in dogs with CME.