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Background: Dromedary camels (Camelus dromedarius) are raised in extremely strict ecological conditions of deserts. Camels are vulnerable to many zoonotic infections. There are limited data on the occurrence of Q fever and borreliosis in camels, in Iran. Aims: The current study was focused on the occurrence of Coxiella burnetii and Borrelia spp. infection in the blood samples of Iranian camels using molecular assays. Effect of the presence of these infections on various hematological factors and some acutephase proteins (Hp, a1AGP, SAA) were also investigated. Methods: Blood samples were collected from 113 clinically healthy camels to investigate the presence of the infections using nested PCR. Moreover, the sequence of positive samples was analyzed phylogenetically. Routine haematological tests were performed and the concentrations of acute-phase proteins were measured in serum using enzyme immunoassay. Results: PCR result showed that 6.19% (95% CI: 2.53-12.35%) (7/113) of camels were positive for C. burnetii. In addition, sequencing results of the corresponding gene of the outer membrane protein (com1) revealed two different genotypes of C. burnetii agent in camels from Southern Iran. In the PCR assay, Borrelia spp. DNA was not detected in the samples. No significant difference was observed in hematological parameters or acute-phase proteins between positive and negative Q fever camels except for mean corpuscular hemoglobin (MCH) and red cell distribution width (RDW). Conclusion: Clinically healthy camels might be very important reservoirs of zoonotic pathogens. Q fever is not considered a notifiable disease in camels of Iran, and clinical cases may scarcely be recognized by the healthcare system. Due to a lack of adequate information, additional studies on the molecular epidemiology and clinical pathology aspects of C. burnetii infection in Iran are needed. [ABSTRACT FROM AUTHOR]

Background: Citral (C10H16O) is the main ingredient of Cymbopogon citratus (lemongrass oil) and can reduce the side effects of oxidative stress. Diabetes caused by insulin deficiency induces oxidative stress in the liver. Aims: This study aimed to investigate the ameliorative effects of citral on selected oxidative parameters and the gene expression of paraoxonase 1 (PON1) and endothelial nitric oxide synthase (eNOS) in a rat model of streptozotocin (STZ)-induced diabetes mellitus. Methods: Forty rats were divided into four groups at random: control (C), control citral (CC), and two STZ-induced diabetic groups (diabetic (D) and citral diabetic (CD)). After diabetes confirmation (day 7), gavage treatment with citral (300 mg/kg body weight (BW)) was started in the CD and CC groups and continued for two weeks. Results: On day 21 of the study, following treatment with citral for 14 days, the serum levels of total antioxidant capacity (TAC), and PON1 in the CD group were significantly increased compared to those in the D group (P<0.05). While treatment with citral caused a significant decrease in the Malondialdehyde (MDA), and eNOS in the CD group compared to those of the D group (P<0.001). The expression rate of liver PON1 gene was considerably upregulated in the CD group compared to that in the D group (P<0.001); while the opposite was observed for eNOS gene expression. However, there was no significant difference between the CC and C groups in terms of all examined parameters (P>0.05). Conclusion: This study showed positive effects of citral on serum antioxidant status and liver gene expression of PON1 and eNOS in diabetic rats. [ABSTRACT FROM AUTHOR]

Abstract: Aims: A variety of pathogens use quorum sensing (QS) to control the expression of their virulence factors. QS interference has hence been proposed as a promising antivirulence strategy. The specific aim of this study was to isolate bacteria from trout tissue able to degrade N‐acyl homoserine lactones (AHL), a QS molecule family. Methods and Results: In total 132 isolates were screened for AHL degradation using Chromobacterium violaceum CV026 as a biosensor. Twenty‐four quorum‐quenching (QQ) isolates were identified biochemically and characterized using 16S rDNA sequencing. They belong to Bacillus, Enterobacter, Citrobacter, Acinetobacter, Agrobacterium, Pseudomonas and Stentrophomonas genera. Four Bacillus spp. showed the highest and fastest QQ activity. AHL degradation proved to be enzymatic in most isolates (except for Stentrophomonas spp. and Pseudomonas sp.) as QQ activity could be destroyed by heat and/or proteinase K treatments. All QQ activity proved to be cell‐bound except for Pseudomonas sp., where it could be detected in the supernatant. The results of aiiA gene homology analysis revealed the presence of aiiA gene encoding AHL lactonase in all examined isolates except Pseudomonas syringae and Enterobacter cloacae. The HXHXDH motif conserved in all AHL lactonases and considered to be essential for AHL degradation was detected in all AiiAs after sequence alignment. Conclusions: Some known and novel QQ bacteria were isolated from trouts and characterized in terms of enzymatic or nonenzymatic AHL degradation activity and their extracellular or intracellular location. In addition, an aiiA gene and its HXHXDH motif were detected in most isolates. Significance and Impact of the Study: We could isolate and identify some novel QQ bacteria including Enterobacter hormaechei, Acinetobacter radioresistens and Citrobacter gillenii. The aiiA gene was detected for the first time in these strains as well as in Stenotrophomonas maltophilia. Our QQ isolates could be used for biocontrol of bacterial infections in aquaculture. [ABSTRACT FROM AUTHOR]

There is little information on the phylogenetic position and life cycle of family Kathlaniidae. Falcaustra araxiana is a member of this family which infects the large intestine of the European pond turtle (Emys orbicularis). In the present study, morphological data and molecular analyses based on the 18S rDNA were performed on different types of F. araxiana originating from the large intestine and gastric nodules in the turtle. Morphological data revealed both larvae and adult stages in the gastric nodules. In addition, all nematodes recovered in the large intestine were adult worms. GenBank accession numbers KM200715 and KM200716 were provided for adult F. araxiana located in the intestine and stomach, respectively, of E. orbicularis. The results of sequencing proved that these two types are completely similar. Accordingly, it can be hypothesized that nodule formation is a part of the life cycle of the parasite or a survival strategy. Furthermore, F. araxiana develops to the adult stage in the gastric mucosa prior to migrating to the large intestine. Phylogenetic analysis revealed that F. araxiana unexpectedly branched away from other members of the superfamily Seuratoidea (Truttaedacnitis truttae, Cucullanus robustus and C. baylisi) and showed a closer relationship with Paraquimperia africana, a member of the Ascaridoidea. It seems that phylogenetic reconstruction for the present parasite needs more detailed morphology, life cycle and molecular studies. [ABSTRACT FROM PUBLISHER]

Hydatidosis is a medically and veterinary important parasitic disease that is endemic in many parts of the world. Unilocular hydatid cysts may develop in almost any part of the body. Up to 70% of hydatid cysts are located in the liver, followed by 25% in the lungs. Cerebral hydatidosis is an uncommon manifestation of the disease, occurring in less than 1/1000 infected hosts, yet diagnosis does pose a problem. We have reported an exceptionally rare case of cerebral hydatidosis in cattle. This is the first report to describe the characteristic pathological features of the cerebral hydatidosis in cattle caused by the G1 genotype of Echinococcus granulosus. Genotypic analysis was performed on a hydatid cyst from a cow originating from southern Iran, based on the sequence analysis of the cox1 mitochondrial gene. [ABSTRACT FROM AUTHOR]

The article discusses a study that characterized the various genotypes of cows based on silent mutation. Study highlights include the impact of bovine pituitary-specific transcription factor 1 variants on milk production, use of Independent Samples Test for comparison of milk production and body condition score between different genotypes, and the importance of thyroid and growth hormones on milk production in cows.

The present study assessed whether the genetic variation among different hosts (sheep and cattle) and geographical isolates (n= 28) of Dicrocoelium dendriticum from Iran is present based on mitochondrial (nad1) and ribosomal (ITS-2) DNA markers. Molecular analysis revealed the presence of at least ten and two distinct haplotypes in the NADH dehydrogenase gene (nad1) and internal transcribed spacer 2 (ITS-2), respectively. The nad1 and ITS-2 sequence data were deposited in GenBank under accession numbers, JX050110–134 and JQ966972–3. According to the results of our study, ND-D and ITS-A are established as being the predominant haplotypes of D. dendriticum in Iran. The Iranian isolates showed a higher intraspecific genetic diversity of 0–0.97% for nad1, compared to 0–0.42% for ITS-2. The alignment and comparison of nad1 and ITS-2 sequences revealed eight and one polymorphic sites, respectively. In the nad1 sequences, six were silent and two nucleotide substitutions were responsible for amino acid alterations. A phylogenetic analysis of the sequence data revealed that host associations and geographic location are not likely useful markers for D. dendriticum haplotype classification. Consequently, sequencing results obtained from the nad1 gene as a mitochondrial marker for the first time in this study would provide a valuable tool to analyse further molecular details of D. dendriticum worldwide. [ABSTRACT FROM PUBLISHER]

Samples from the kidney of 100 diseased fish with signs of haemorrhagic septicemia and 50 samples from outlet water of fish farms were taken aseptically and cultured. In the laboratory, 75 colonies of gram-negative bacteria were biochemically diagnosed as motile aeromonads in our Bacteriology Laboratory Unit using API 20E rapid identification system. The genotype identification using specific primers for 16S rDNA by PCR and direct sequencing of 28 Iranian motile aeromonads isolates were as follow: in diseased fish, Aeromonas hydrophila (3 isolates, 15%), A. veronii bv. sobria (8 isolates, 40%), A. bestiarum/piscicola (5 isolates, 25%), A. media (3 isolates, 15%), A. jandaei (1 isolate, 5%), A. aquariorum (0 isolate, 0%) and in water, Aeromonas hydrophila (0 isolate, 0%), A. veronii bv. sobria (6 isolates, 75%), A. bestiarum/piscicola (0 isolate, 0%), A. media (1 isolate, 12.5%), A. jandaei (0 isolate, 0%) and A. aquariorum (1 isolate, 12.5%). Results of this study suggest that the incidence of motile aeromonads septicemia due to A. veroniibv. sobriais the most prevalent motile aeromonads. Nucleotide polymorphisms on the sequencing results of the 16S rDNA were detected as noticeable inter and intra-specific variation within the population of different aeromonads isolates. In total, 10-20 variant nucleotide positions in this region were observed among Aeromonas spp. [ABSTRACT FROM AUTHOR]

Abstract: Objective: To investigate the relationship between polymorphism within the 5′-untranslated region (5′-UTR) of IGF-I gene and its periparturient concentration in Iranian Holstein dairy cows. Methods: Blood samples (5 mL, n = 37) were collected by caudal venipuncture from each animal into sample tubes containing the EDTA and DNA was extracted from blood. In order to measure IGF-I concentration the collection of blood samples (n = 111) was also done at 14 d before calving (prepartum), 25 and 45 d postpartum. Results: We found evidence for a significant effect of C to T mutation in position 512 of IGF-I gene on its serum concentration in dairy cows in Iran. Cows with CC genotype had significantly higher concentration (Mean±SD) of IGF-I at 14 d prepartum (91.8±18.1) μg/L compared to those with TT genotype (73.3±14.4) μg/L (P=0.04). A significant trend (quadratic) was found for IGF-I concentration, as higher in CC cows compared to ones with TT genotype, during the 14 d before calving to 45 d postpartum (P=0.01). Conclusion: We concluded that C/T transition in the promoter region of IGF-I gene can influence the serum concentration of IGF-I in periparturient dairy cows. [Copyright &y& Elsevier]

Background: Dogs may be infected with species of Leishmania parasites that are disseminated through blood circulation and invade the internal organs. In this study, we aim to detect the parasite in the blood of dogs using the PCR technique. The present work was performed from February 2022 to May 2023 in Fars Province, southern Iran, where the disease is endemic., Results: In total, 7(5.1%) out of 135 blood samples, six were identified as Leishmania tropica and one as Leishmania major. We found no trace of Leishmania infantum, which is always known for visceral infection. In addition, no sign of cutaneous lesions or a significant disease was seen in the animals infected with both species. Of 48 dogs with anemia, two were Leishmania positive. The mean value of hematological parameters in the infected dogs was within the normal range except for a significant reduction in the platelet measures (p < 0.05)., Conclusions: Our data revealed that both Leishmania species, tropica and major, may manifest as viscerotropic leishmaniasis. More investigations are needed to understand the conditions under which these species choose the type of infection. Moreover, our data emphasize the role of asymptomatic dogs in carrying these parasites, a crucial factor in spreading the disease., (© 2024. The Author(s).)

Aim: Tempol, a synthetic antioxidant compound, has received significant attention for its potential therapeutic applications in recent years, especially against ischemia/reperfusion (I/R) injury. The aim of the present research was to assess the protective effects of Tempol on testicular I/R injury caused by testicular torsion and detorsion (T/D) in rats., Methods: The subjects were divided into five groups: sham, testicular T/D, testicular T/D with Tempol treatment at 50 and 100 mg/kg, and healthy rats treated with Tempol at 100 mg/kg. Testicular torsion was induced by rotating the left testicles for 2 h, followed by detorsion for 24 h. Testicular tissues were evaluated for gene expression, oxidative stress markers, and histopathology, epididymal sperms were stained and analyzed, and blood serum samples were collected to measure the testosterone hormone., Results: The results showed that testicular I/R caused a significant decrease in sperm velocity parameters, viability, and count, as well as an increase in abnormal sperms (p < 0.05). However, treatment with Tempol significantly improved these parameters (p < 0.05). Histopathological analysis revealed severe damage to the testicular tissues, but treatment with Tempol improved the structural integrity of the seminiferous tubules. Testicular I/R also resulted in increased oxidative stress index and decreased testosterone levels significantly (p < 0.05), but Tempol administration mitigated these effects significantly (p < 0.05). Furthermore, the expression of Bax and Bcl 2 , genes associated with apoptosis, were significantly altered by testicular I/R (p < 0.05), but Tempol prevented these changes significantly (p < 0.05)., Conclusion: These findings provide strong evidence that Tempol can effectively prevent testicular I/R injury., Competing Interests: Declaration of competing interest None., (Copyright © 2024. Published by Elsevier Inc.)

Background: Parasitic infections pose significant threats to humans' and animals' well-being worldwide. Among these parasites, Halicephalobus spp., a genus of nematodes, has gained attention due to its ability to cause severe infections in various animal species, including horses., Objective: This study aimed to determine the prevalence of Halicephalobus spp., specifically focusing on Halicephalobus gingivalis in horses., Materials and Methods: In July 2022, a cross-sectional study was conducted in northern Iran to determine the prevalence of Halicephalobus spp. Using standard coprological techniques, 141 fecal samples from randomly selected horses were analyzed for GI helminth eggs. The Halicephalobus spp. eggs present in faeces were identified by molecular methods. Polymerase Chain Reaction (PCR) was used to amplify the partial 5' variable region (~ 390 base pairs) of 18 S DNA using SSUA&#95;F and SSU22&#95;R primers. Furthermore, the PCR products obtained were sequenced, and phylogenetic analysis was performed using available sequences from GenBank., Results: Microscopic examination of 141 fresh faecal samples revealed 5 fecal samples were infected with small ellipsoidal nematode eggs ranging between 40 and 50 × 50-60 μm. This study's PCR amplicons showed ~ 390 bp bands on 2.0% agarose gel. A partial sequence of 18 S DNA (363 bp) was obtained herein (GenBank accession no. OQ843456)., Conclusion: Overall, using molecular tools represents a significant step forward in diagnosing and managing the Halicephalobus gingivalis infections in horses., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Trypanosoma species cause animal trypanosomiasis that infects many animals. Trypanosoma evansi is an organism that infects camels. There are many economic problems associated with this disease, including lower milk and meat yields and abortions. The purpose of the current survey was molecular study of the presence of Trypanosoma in dromedary camel blood in the south of Iran, and its effects on the hematologic, and some acute-phase protein changes. Blood samples were aseptically collected from the jugular vein of dromedary camels (n = 100; aged from 1 to 6 years) originating from Fars Province in EDTA-coated vacutainers. Genomic DNA from 100 µL of the whole blood was extracted and amplified using a PCR assay based on ITS1, 5.8S, and ITS2 ribosomal regions. Also, the PCR products obtained were sequenced. Moreover, the changes in hematological parameters and serum acute-phase proteins (serum amyloid A, alpha-1 acid glycoprotein, and haptoglobin) were measured. Among 100 tested blood, nine samples (9%, 95% CI: 4.2-16.4%) were found positive by the PCR assay. The phylogenetic tree and blast analysis showed four different genotypes closely related to the strains (accession numbers: JN896754 and JN896755) previously reported from dromedary camels in Yazd Province, center Iran. Based on hematological analysis, normocytic and normochromic anemia and lymphocytosis were detected in the PCR-positive cases compared with the negative group. Furthermore, alpha-1 acid glycoprotein was significantly increased in the positive cases. There was a substantial and positive relation between the number of lymphocytes, and the levels of alpha-1 acid glycoprotein and serum amyloid A in the blood (p = 0.045, r = 0.223 and p = 0.036, r = 0.234, respectively). A noticeable frequency of T. evansi infection was reported in dromedary camels in south Iran. This is the first report on the genetic diversity of T. evansi in this region. There was a significant association among Trypanosoma infection, lymphocytosis, and alpha-1 acid glycoprotein. Trypanosoma-positive camels had a significant decrease in hematocrit (HCT), hemoglobin (Hb), and red blood cell (RBC) values compared to the non-infected group. Further experimental studies are needed to elucidate the hematological and acute-phase protein alteration during a different phase of Trypanosoma spp. infection., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase), as a key factor in innate immunity, consists of several components, one of them is p40phox which is encoded by neutrophil cytosolic factor 4 (NCF4). Respiratory burst and reactive oxygen species (ROS) production are antimicrobial mechanisms associated with NADPH oxidase. This study evaluated the effects of g.18174 A > G and g.18270C > T single-nucleotide polymorphisms (SNP) in NCF4 on bovine mastitis and the respiratory burst capacity of neutrophils. SNPs of 160 dairy cattle were determined using a novel PCR-RFLP protocol by employing restriction enzymes, MboI and FokI. Also, the flow cytometry measured respiratory burst in 82 blood samples. Our results indicated that only g.18174 A > G SNP reduced the respiratory burst capacity. However, both SNPs were not significantly correlated with clinical mastitis. We concluded that g.18174 A > G decreases the function of NADPH oxidase. However, both SNPs were not significantly correlated with clinical mastitis., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest. Furthermore, the authors declare that the research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Canine hepatozoonosis is a tick-transmitted apicomplexan infection caused by two species of Hepatozoon, H. canis , and H. americanum . The present research aimed at detection of Hepatozoon spp. in dogs and its effects on hematological alterations. Blood samples were taken from 108 dogs to assess Hepatozoon spp. Phylogenetic analysis was performed based on the 18S rDNA marker by PCR assay and Giemsa-stained blood smear examination. Of the 108 blood samples of dogs tested in the present study, eight (7.40%, 95% CI: 3.25-14.07%) were positive by the Hepatozoon-specific PCR assay. However, in the microscopic examination, only one sample (0.93%) was positive. All of the sequenced samples were H. canis. The Hepatozoon sequences obtained from PCR amplicons in the canine-positive cases exhibited 100% similarity to each other and 98.47-100% similarity to other relevant sequences in GenBank. These findings represent the first molecular evidence of H. canis in dog populations in South Iran. Furthermore, according to the hematological analysis, significantly higher average numbers of neutrophils and lymphocytes were found in the infected group compared to the non-infected dogs. In this study, no statistically significant connection (P<0.05) was observed between H. canis infection and the examined risk factors., Competing Interests: The authors declare that they have no conflicts of interest.

In all equids worldwide, Theileria equi and Babesia caballi are believed to be two important erythrocytic protozoa that cause equine piroplasmosis. In addition, it was recently discovered that Theileria haneyi is another potential equine piroplasmosis (EP) agent. Ixodid ticks are the major vectors of these parasites. Equine piroplasmosis is of international importance and affects enormously the equine industry. In this study, for the first time, molecular prevalence and genetic diversity of piroplasma parasites (T. equi and B. caballi) in horses from Fars province (south of Iran) were determined. Also, hematological alterations of naturally infected horses were analyzed. PCR positive horses showed anemia, thrombocytopenia, leukocytosis with a left shift of neutrophilia, and monocytosis. PCR results revealed that, from 133 blood samples of horses, 40 samples were positive (30.07%). The occurrence of T. equi in this area (30.07%) was more than the national average prevalence of T. equi (24.11%), but B. caballi prevalence in study area (0%) was less than the average of previous studies in Iran (5.47%). Our findings revealed that the T. equi was widespread in Fars province of Iran. PCR products of 18S rDNA and EMA-1 genes of T. equi strains were sequenced successfully. All 18S rDNA sequences collected in this experiment revealed 100% similarity together. According to the phylogenetic tree constructed using the 18S rDNA gene, Iranian T. equi is clustered with strains from Cuba (KY111762, KY111761) and USA (CP001669, JX177672). So, this could be concluded that T. equi studied in this research, and those strains are initiated from a common T. equi ancestor at an unknown time ago. Also, the phylogenetic tree based on EMA-1 gene demonstrated a genetically diverse population of Iranian T. equi strains (10 different genotypes). As EMA-1 is one of the most immunogenic antigens in this parasite, such variability could be a concern about the efficacy of T. equi vaccines. Finally, more studies on equine piroplasmosis in the provinces of the southern region of Iran are recommended to create a better vision of disease in this region., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The blood serum of dromedary camels contains a unique type of antibodies with a high potency to neutralize toxins and to identify and inactivate some bacterial pathogens. The present study was designed to examine changes in the endometrial histology of cows with no subclinical endometritis (SE) (experiment 1) and changes in the uterine cytology and endometrial mRNA expression of COX2, IL-1β, IL-8, and iNOS following intrauterine administration of DCBS in cows with SE as compared to different common treatments (experiment 2). In addition, the effects of the intrauterine administration of DCBS were examined on the pregnancy rate in dairy cows with SE (experiment 3). DCBS did not induce any histological reactions in the bovine endometrium. The mean ( ± SE) percentage of PMNs after intrauterine infusion of Pen-Strep, DCBS and double DCBS in cows with SE differed as compared to cows treated with PGF2α and no treated cows with SE (1.47 ± 0.87; 1.43 ± 1.08 and 1.31 ± 0.23 vs 3.00 ± 0.43 and 3.5 ± 0.75, P < 0.05, respectively) in experiment 2. The mRNA expression of COX2, IL-1β, and iNOS was reduced (P < 0.05) after treatment with Pen-Strep, DCBS and double DCBS as compared with no treated-cows with SE. The pregnancy rate after the first AI was tended to be higher (49.2 vs 39.0%), while the overall pregnancy rate was greater (P < 0.05) in cows with SE when treated with DCBS as compared to the Pen-Strep group (76.9 vs 61.0%) in experiment 3. In conclusion, serum of dromedary camel, as a non-antibiotic preparation, can improve the uterine health and fertility when used for the treatment of bovine SE., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The objective was to investigate the multidrug resistance and presence of class 1 and 2 integrons in 300 Escherichia coli isolates obtained from 20 broiler farms during three rearing periods (one-day-old chicks, thirty-day-old chickens, and one day before slaughter) in Fars, South Iran. Results showed that 81.00%, 82.00%, and 85.00% of isolates were multidrug-resistant on the first day, thirty-day-old chickens, and one day before slaughter, respectively. Multidrug-resistant E. coli isolates were further examined for the presence of class 1 and 2 integrons using PCR assay. The existence of class 1 integron-integrase gene ( intI1 ) was confirmed in 68.40%, 72.70%, and 60.90% of multidrug-resistant isolates from stage 1, stage 2, and stage 3 of the rearing period, respectively. The frequency of class 2 integron-integrase gene ( intI2 ) during the first to the third stage of sampling was 2.60%, 25.50%, and 30.40%. Also, sequence analysis of the cassette arrays within class 1 integron revealed the presence of the genes associated with resistance for trimethoprim ( dfr A), streptomycin ( aad A), erythromycin ( ere A), and orf F genes. The results revealed that percentages of antimicrobial resistance in E. coli isolates were significantly higher in the middle and end stages of the rearing period. In conclusion, widespread dissemination of class 1 integrons in all three stages and rising trends of class 2 integrons existence in E. coli isolates during the rearing period of broiler chickens could exacerbate the spread of resistance factors among bacteria in the poultry industry. Future research is needed to clarify its implication for human health., (© 2021 Urmia University. All rights reserved.)

Oxidative stress, caused by extreme accumulation of un-scavenged reactive oxygen species, plays an integral role in the Ischemia-Reperfusion (I/R) injury to the testicles following testicular torsion. The current research aimed to examine the protective effects of crocin as a natural antioxidant on testicular I/R injury in rats. Animals were divided randomly into five groups (seven each): (1) sham group, (2) torsion/detorsion (T/D) group, (3) intact group with 100 mg/kg crocin, (4) and (5) T/D groups followed by treatment with two different doses of crocin (50 and 100 mg/kg (IP)). I/R injury was induced by 720° clockwise torsion of the left testicles for 2 h. After 24 h of reperfusion, blood samples and epididymal sperms were collected to measure biochemical (GPx, SOD, and MDA), hormonal (testosterone), and sperm parameters (total sperm recovery, motility, viability, and morphology). Moreover, affected testicles were subjected to histopathology examination. I/R injury caused a significant reduction in sperm characteristics (except for morphology) (P < 0.05), which could not be significantly improved by crocin administration at either dose (P > 0.05). Johnsen's testicular score, mean seminiferous tubular diameter, and germinal epithelial cell thickness were significantly decreased in the T/D group compared to the intact and sham groups. However, crocin could significantly improve the histopathological parameters in both treatment groups compared to the T/D group (P < 0.05). T/D reduced SOD and GPx activity and testosterone level significantly (except for GPx) compared to the sham group (P < 0.05). However, crocin administration could significantly reverse them. Also, crocin reduced the amount of MDA significantly in the high-dose treatment group in comparison to T/D group (P < 0.05). The results of the current study revealed that crocin could be a promising agent to protect against I/R injury following surgical correction of the testicular torsion., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2021 Elsevier Inc. All rights reserved.)

In this study, the relationships of integron 1 element, formaldehyde dehydrogenase, and orfF genes with the level of formaldehyde resistance of isolated E. coli were investigated. E. coli bacteria were isolated from apparently healthy and colibacillosis-affected broilers of Fars Province, Iran. Formaldehyde resistance level and the presence of genetic markers were measured using MIC, and PCR tests, respectively. The prevalence of integron 1 element, orfF, and formaldehyde dehydrogenase genes in E. coli isolates were 61%, 8%, and 94%, respectively. In addition, according to our cut off definition, 15% and 85% of isolates were resistant and sensitive to formaldehyde, respectively. None of the genes had a statistically significant relationship with the formaldehyde resistance; however, the isolates containing integron 1 were significantly more sensitive to formaldehyde in the MIC test than those without integron 1. Integron 1 gene cassette could carry some bacterial surface proteins and porins with different roles in bacterial cells. Formaldehyde could also interfere with the protein functions by alkylating and cross-linking, and this compound would affect bacterial cell surface proteins in advance. Through an increase in the cell surface proteins, the presence of integron 1 gene cassette might make E. coli more sensitive to formaldehyde. As integron 1 was always involved in increasing bacterial resistance to antibiotics and disinfectants such as QACs, this is the first report of bacterial induction of sensitivity to a disinfectant through integron 1. Finally, integron 1 does not always add an advantage to E. coli bacteria, and it could be assumed as a cause of vulnerability to formaldehyde., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Mugger crocodile is the only crocodile existing in Iran. The present study was aimed to investigate the bacterial flora in oral and cloacal cavities of wild Mugger crocodiles in Negour protected area, Iran. The isolation and molecular characterization of oral and cloacal bacterial flora were performed in 22 Mugger crocodiles captured in Negour protected area, Iran. Ten bacterial species from all oral samples and six bacterial species from all cloacal samples were recovered. The most commonly isolated bacteria in oral samples were Burkholderia contaminans and Lactococcus garvieae , respectively; whereas, in cloacal samples, it was Lactococcus lactis . It is likely that the isolated bacteria would pose a threat to both crocodiles and humans health. It can threaten crocodiles during stressful conditions; while, humans would be susceptible if they are bitten by crocodiles, consume their meat or spend time near their natural environment. This study provides useful information about bacterial diversity which could help to select the most appropriate anti-bacterial when dealing with infections caused by crocodiles., Competing Interests: The authors declare that there is no conflict of interest.

Background: Lactococcus garvieae causes lactococcosis in rainbow trout in many parts of the world., Aims: This study was conducted for the existence of the virulent factors and differentiation of the two serotypes in L. garvieae ., Methods: Twenty-two strains of L. garvieae isolated from diseased rainbow trout from farms in different regions and mugger crocodile of Iran, were investigated. In order to rapidly detect the presence of the hly1 , hly2 , hly3 , NADH oxidase , sod , pgm , adhPsaA , eno , LPxTG-3 , adhCI , and adhCII virulence genes, two multiplex polymerase chain reaction (PCR) assays were developed. Also, simplex PCR method was used to identify the bacterial serotypes, CGC , LPxTG-2 , Adhesion , and adhPav virulence genes using the specific primer., Results: All varieties of L. garvieae contained the hly1 , hly2 , hly3 , NADH oxidase , pgm , adhPav , LPxTG-3 , sod , eno , adhPsaA , adhCI , and CGC virulence genes. Also, adhCII gene was present in all strains except one of the isolates originated from mugger crocodile. In addition, LPxTG-2 gene was only present in one of the isolates belonging to mugger crocodile. Adhesion gene was not present in all the strains. Interestingly, all the 22 strains originated from both hosts were identified as belonging to the serotype I. Based on the phylogenetic sequences of the capsule gene cluster, group all fish isolates into a cluster together with one isolate obtained from mugger crocodile., Conclusion: Further studies are recommended to investigate the role of virulence genes in L. garvieae and evaluate their pathogenicity to rainbow trout.

Background: Fasciola hepatica as an important parasite affects health of humans and animals in some tropical and subtropical areas of the world, including Iran. Little is known about the molecular diversity of Fasciola in Equidae. Therefore, this study aimed to characterize the genetic polymorphisms among parasites., Methods: Eight adult Fasciola spp. isolates were collected from a working donkey after necropsy in Shiraz, southwestern Iran, in 2018. Primarily, various parameters were measured morphologically. Subsequently, DNA was extracted from each fluke and molecular markers of cytochrome C oxidase ( cox 1) and NADH dehydrogenase 1( nad 1) from individual Fasciola isolates were amplified using PCR assay and sequence data were employed for molecular and phylogenetic analysis. Genetic diversity between isolates was evaluated by comparing the sequences of these two mitochondrial regions., Results: Based on the morphological and analyzed mitochondrial sequences, all of eight donkey isolates (100%) were identified as F. hepatica . Moreover, nine and five nucleotide polymorphisms were identified in the cox 1and nad 1 region sequences, respectively., Conclusion: Accordingly, phylogenetic data revealed five and four haplotypes among donkey isolates based on the cox 1and nad 1 markers. Similarly, some of these haplotypes have been previously reported from different host species in Iran as well as all around the world., Competing Interests: Conflict of interest The authors declare that they have no conflict of interest., (Copyright © 2020 Asadpour et al. Published by Tehran University of Medical Sciences.)

Emerging Bartonella spp. infection can result in clinical symptoms such as endocarditis in humans and animals. This study analyzed the genetic phylogeny of the Bartonella spp. circulating in Iranian dogs. Also, this is first study on the relationship of Bartonella spp. and haematological factors from dogs in Fars. Ninety-eight blood samples were collected from the dogs of Fars province, Iran. Two different PCRs targeting rpoB gene and ITS sequence of Bartonella spp., followed by sequencing were performed. In addition, CBC and the differential count of WBC were determined. The "prevalence" of Bartonella spp. was 12.2 % (95 % CI: 5.72-18.68 %) in this population and the sequences matched with a newly proposed species; 'Candidatus Bartonella merieuxii'. A significant increase in WBC due to neutrophilia and decreased RBC, Hct, and Hb concentrations were detected in Bartonella spp. infected dogs. The close contact between humans and dogs, and the zoonosis potential of Candidatus Bartonella merieuxii, emphasize on the need for more studies on 'Candidatus Bartonella merieuxii'., (Copyright © 2020. Published by Elsevier Ltd.)

Hemotropic mycoplasmas (haemoplasmas) are wall-less bacteria, which may lead to anemia and, even, mortality in mammals. The present study was aimed to characterize the causative agent(s) of haemoplasma infection in blood samples taken from horses (n = 133) in south of Iran. Microscopic examination of blood smears and PCR assay were performed for the detection of hemotropic Mycoplasma and equine piroplasma (Babesia caballi and Theileria equi). For the purpose of molecular characterization, 16S rDNA and 18S rDNA markers were used for hemotropic Mycoplasma and piroplasma pathogens, respectively. The PCR-positive samples were sequenced for haemoplasma and further phylogenetic analysis was performed for the obtained haemoplasma sequences. Nine out of 133 (6.77 %, 95 % CI: 2.5-11.04 %) horses were positive for Mycoplasma sp. by PCR. Furthermore, three of these animals were co-infected with T. equi. Interestingly, the phylogenetic and molecular analysis of the haemoplasma sequences derived from the PCR amplicons in the equine positive cases showed 100 % identity with 16S rDNA in Mycoplasma ovis, an HM member mainly found in sheep and goats. In addition, the hematological analysis showed that PCR positive horses for M. ovis-like species had a lower hematocrit (Hct), hemoglobin (Hb) concentration, and red blood cell (RBC) count compared to the PCR negative ones (P ≤ 0.05). Mild anemia was also developed in the haemoplasma-positive horses. These findings represent the first molecular evidence of M. ovis-like species in horses. Further experimental studies are needed to examine the importance of this nonspecific host infection and evaluate its pathogenicity in equine and other species., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Aeromonas spp. is one of the major pathogens of freshwater animals. There has been little research on the genetics of antimicrobial resistance associated with it in Iranian aquaculture. To remedy this lack in research, 74 multi-drug-resistant Aeromonas spp. were isolated from farmed diseased carp, trout, sturgeon, ornamental fish, crayfish, and corresponding water samples and examined for genomic integron sequences. Class 1 integrons, containing seven types of integron cassette arrays ( dfrA1-aadA1 , dfrA1-orfC , dfrA12-aadA2 , dfrA12-orfF-aadA2 , dfrA15 , dfrB4-catB3-aadA1 , aac(6')-Ib-cr-arr3-dfrA27 ) were found in 15% of the resistant isolates; no class 2 integrons were detected in any of the resistant isolates. As some tested isolates were resistant to more than two groups of antibiotics, our results demonstrated that freshwater animals in Iran could be a source of multiply drug-resistant Aeromonas spp. This finding suggests that the origin of the antimicrobial resistance of these animals be placed under increased surveillance in the future and that the use of antimicrobials be limited in aquaculture.

The genus Bartonella is comprised of Gram-negative coccobacilli, aerobic, and facultative intracellular bacteria which are transmitted by hematophagous vectors (e.g., fleas, lice, sandflies, and ticks). Each species of Bartonella infects one or few related mammals as reservoir host(s). If a Bartonella spp. infects a nonspecific host like humans, it can lead to a more acute disease. Bartonella spp. has been detected more recently for the first time in camels in Israel by Rasis and colleagues. However, the epidemiological and public health importance of this new pathogen in camels is not clear. In this study, we aimed to detect the Bartonella spp. in the blood samples of Iranian camels, measure their prevalence, and determine their species. Also, the relationship between Bartonella spp. infection and different hematological factors and acute-phase proteins (Hp, a1AGP, SAA) was investigated. Finally, the sequences of three DNA regions, i.e.16S rDNA, rpoB, and ITS, were determined and phylogenetically analyzed. From the 106 examined blood samples of camels from Fars province (southern area of Iran), 18 samples were positive (17%). The findings also showed that Bartonella spp. positive camels had significantly lower Hb, MCH, and MCHC but higher RDW, SAA, and WBC (P < 0.05) compared to the control group. Our Bartonella strain was genetically similar to the 'Candidatus Bartonella dromedarii' but different from Bartonella bovis. Thus, more studies are required to investigate the phenotypic and genotypic characteristics of 'Candidatus Bartonella dromedarii'. Also, there is a need to evaluate precisely the risk factors, transmission routes, and zoonotic potential of this species., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In this study, we evaluated the impact of the catecholamines on growth, swimming motility, biofilm formation and some virulence factors activities of pathogenic Yersinia ruckeri. Norepinephrine and dopamine (at 100 µM) significantly increased the growth of Y. ruckeri in culture media containing serum. An increase in swimming motility of the pathogen was found following the exposure to the hormones; however, no effect was seen on caseinase, phospholipase and haemolysin productions. Further, antagonists for the catecholamine receptors were observed to block some of the influences of the catecholamines. Indeed, the effects of catecholamines were inhibited by chlorpromazine (the dopaminergic receptor antagonist) for dopamine, labetalol (α-and β-adrenergic receptor antagonist) and phenoxybenzamine (the α-adrenergic receptor antagonist) for norepinephrine, but propranolol (the β-adrenergic receptor antagonist) showed no effect. Pretreatment of Y. ruckeri with the catecholamines resulted in a significant enhancement of its virulence towards rainbow trout and the antagonists could neutralize the effect of the stress hormones in vivo. In summary, our results show that the catecholamines increase the virulence of Y. ruckeri which is pathogenic to trout through increasing the motility, biofilm formation and growth., (© 2019 John Wiley & Sons Ltd.)

Co-infections occur when a host is infected by 2 or more different pathogen types, either by secondary or simultaneous infections, and are very frequent in nature. In this study, 10 narrow-clawed crayfish Astacus leptodactylus with signs of disease were collected from Haft Baram Lake (Fars province, southern Iran). Samples of fluid from inside the intact abscess and melanized lesions in the cuticle were cultured aseptically onto brain heart infusion agar and Sabouraud dextrose agar for bacterial and fungal agents, respectively. After primary colony isolation for bacterial and fungal agents, the isolates were confirmed as Aeromonas hydrophila and Fusarium solani, using specific PCR methods based on 16S rDNA and internal transcribed spacer (ITS) rDNA sequences that produced a single band of 685 bp and 600 bp, respectively. Partial sequence analysis of the F. solani ITS showed 100% sequence identity among all our samples, as well as a close genetic relationship between this isolate (GenBank accession number MG519784) and those previously reported from loggerhead sea turtle Caretta caretta in Cape Verde (FJ948133, AM412641, and DQ535186), black-blotched stingray Taeniura melanopsila in Japan (LC019016), and American manatee Trichechus manatus in Japan (AB775569). The results indicate that narrow-clawed crayfish can be infected by A. hydrophila and F. solani simultaneously, and to the best of our knowledge, this is the first report of just such a co-infection in this host. Further studies are necessary to investigate the pathogenicity of these organisms and their effects on narrow-clawed crayfish.

Five N-acyl homoserine lactone-degrading bacteria (quorum quenching (QQ) strains) were selected to evaluate their impacts on growth, virulence factors and biofilm formation in Yersinia ruckeri in vitro. No difference was observed among the growth pattern of Y. ruckeri in monoculture and coculture with the QQ strains. To investigate the regulation of virulence factors by quorum sensing in Y. ruckeri, cultures were supplemented with 3oxo-C8-HSL. The results indicated that swimming motility and biofilm formation are positively regulated by QS (p < 0.05), whereas caseinase, phospholipase and haemolysin productions are not influenced by 3oxo-C8-HSL (p > 0.05). The QQs were able to decrease swimming motility and biofilm formation in Y. ruckeri. QQ bacteria were supplemented to trout feed at 10 8  CFU/g (for 40 days). Their probiotic effect was verified by Y. ruckeri challenge either by immersion or injection in trout. All strains could significantly increase fish survival with Bacillus thuringiensis and Citrobacter gillenii showing the highest and lowest relative percentage survival (RPS) values (respectively, 85% and 38%). Besides, there was no difference between the RPS values by either immersion or injection challenge expect for B. thuringiensis. The putative involvement of the QQ capacity in the protection against Yersinia is discussed., (© 2018 John Wiley & Sons Ltd.)

The purpose of the present study was to isolate and identify the pathogenic agents in Acipenser stellatus (Pallas, 1771) and Huso huso, (Linnaeus, 1758) reared in the south of Fars province, Iran which have shown infectious disease signs. Samples from spleen and kidney of 32 fishes showing septicemia symptoms such as decreasing of appetite, unbalanced swimming, expanded wounds, and petechia on the body surfaces, pectoral fins rot, visceral hemorrhage, bleeding on the spleen, and heart ascites were collected. Then samples were cultured on brain heart infusion agar growth media, stain and biological and biochemical tests on purified bacteria were performed. On the other hand, 16S rDNA region of the isolated organism was amplified using PCR. The amplified gene fragment was sequenced and evolutionary history was inferred by phylogenetic tree construction using neighbor-joining method. Results indicated that two bacterial species including Chryseobacterium joostei which isolated from the kidney of stellate sturgeon (43.00%), and Aeromonas veronii which isolated from the spleen of both sturgeon species (75.00% and 31.00% from beluga and stellate sturgeon, respectively), were recognized. Phylogenetic tree analysis showed that Fars isolated organisms including A. veronii and C. joostei had highest similarity with A. veronii bv veronii and C. joostei isolated from France, respectively., Competing Interests: The authors of the paper do not declare a competitive financial interest.

Follicle stimulating hormone (FSH) is released from the anterior pituitary gland and has an important role in female fertility. As FSH is a glycoprotein polypeptide hormone which cannot pass through the cell membrane, its influence on target cells must be mediated by the FSH receptor (FSHR). Accordingly, any kind of mutation in FSHR can affect reproduction in dairy cows. In this study, the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used for recognition of a point mutation (A/G: position -278) located in the FSHR gene in Iranian dairy cows. The association was evaluated of this mutation with reproductive performance. Blood samples were collected from 79 cows in a dairy farm in Iran and genotyped based on this single nucleotide polymorphism (SNP). The 5'-flanking regions of FSHR gene were successfully amplified and produced a fragment of 211bp in all cases. Three different patterns were, however, produced following restriction digestion with FaqI enzyme. The molecular results showed the existence of three different genotypes of AA, AG and GG among examined cows. In this study percentages of genotypes were 51.9%, 43.2% and 4.9% for AA, AG and GG genotypes, respectively. Allele frequencies were 73.5% and 26.5% for A and G, respectively. Results indicate that cows lacking allele G had desirable fertility in which a greater percentage (53.7%) of cows lacking Allele G (AA) had services per conception (SPC) of <2 in the previous lactation; while a lesser percentage of cows with Allele G (28.9%) had SPC of <2 (P<0.05). There was no difference in the days non-pregnant (DNP) and calving to first service interval among cows with these genotypes (P>0.05). Calving to first service interval was 69.9 ±12.3 in cows with Allele G and 74.73±13.9 in cows without Allele G (P>0.05). Percentage of cows with repeat breeder syndrome (SPC >3) was also 15.6% and 27.6% in cows without Allele G and with Allele G, respectively, but these values were not different (P>0.05). It can be concluded that the A to G mutation within the upstream region of FSHR gene (position -278) may affect some reproductive variables in Holstein dairy cows., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Dactylogyrids are a group of monogenean parasites that have a high species intensity on the gills of cyprinid fish. In this study, 89 common carp (Cyprinus carpio) and 25 grass carp (Ctenopharyngodon idella) were collected from Mashhad, in northeastern Iran. A total of 31 Dactylogyrus specimens, 20 and 11 specimens from the gills of common carp and grass carp, respectively, were collected and studied by morphologic analysis and molecular analysis based on 28S rDNA. Four lineages were revealed: D. anchoratus, D. extensus, D. lamellatus, and a new species. Phylogenetic analysis showed that 2 species, namely D. extensus and D. lamellatus, were identical to those previously reported. In addition, nucleotide sequencing showed the greatest homology (93.01%) of D. anchoratus to be with a species registered as D. inexpectatus in the Basic Local Alignment Search Tool database. The new Dactylogyrus species formed a distinct clade of its own in the phylogram. From the morphologic, molecular, and phylogenetic evidence, we propose that this isolate is a single new species within the genus Dactylogyrus. Further phylogenetic analysis, however, including the incorporation of additional molecular targets, is required to infer relationships among species in the Dactylogyrus genus.

Consumption of poultry products contaminated with Salmonella is one of the major causes of foodborne diseases worldwide and therefore detection and differentiation of Salmonella spp. in poultry is important. In this study, oligonucleotide primers were designed from hemD gene and a PCR followed by high-resolution melt (HRM) curve analysis was developed for rapid differentiation of Salmonella isolates. Amplicons of 228 bp were generated from 16 different Salmonella reference strains and from 65 clinical field isolates mainly from poultry farms. HRM curve analysis of the amplicons differentiated Salmonella isolates and analysis of the nucleotide sequence of the amplicons from selected isolates revealed that each melting curve profile was related to a unique DNA sequence. The relationship between reference strains and tested specimens was also evaluated using a mathematical model without visual interpretation of HRM curves. In addition, the potential of the PCR-HRM curve analysis was evaluated for genotyping of additional Salmonella isolates from different avian species. The findings indicate that PCR followed by HRM curve analysis provides a rapid and robust technique for genotyping of Salmonella isolates to determine the serovar/serotype.

Ehrlichiosis is a zoonotic disease which has been reported from some regions of Iran. This study was aimed to determine the presence and prevalence of ehrlichiosis in suspected dogs referred to the Faculty of Veterinary Medicine, Shiraz University, Shiraz, Iran using polymerase chain reaction (PCR). Blood samples were collected from 98 suspected dogs with at least one of the five following findings: thrombocytopenia, anemia (hematocrit < 37.00%), gastrointestinal signs and respiratory and/or central nervous system diseases. Complete blood count was performed for each sample. After genomic DNA extraction, PCR assay was carried out using a commercial PCR kit. The results showed that only three out of 98 samples (3.06%) were positive for ehrlichiosis. There was no significant difference in hematological parameters between infected and non-infected cases. These results emphasize that ehrlichiosis has a low prevalence among examined cases in southern Iran. Further serological and molecular studies are needed to clarify the epidemiological feature of this infection in different areas of Iran.

To date, a number of species of Haemogregarina have been described from different turtle hosts, mainly based on the morphology of the developmental stages detected in the host erythrocytes. The diversity and overlapping morphological features in the old and recent descriptions has led to considerable complications in the taxonomy of Haemogregarina spp. In this study, different stages of maturity and developing gamonts of a putative new species of Haemogregarina were detected in erythrocytes of the Caspian turtle Mauremys caspica (Gmelin) (Geoemydidae) originating from a southern province in Iran. Although some of the morphological characteristics were consistent with Haemogregarina stepanowi Danilewsky, 1885, some new observations were made, particularly in the gamont stage. The phylogenetic analysis based on 18S rDNA sequences revealed that the present isolate appears as basal to a large clade of Haemogregarina spp. with sequences available in the GenBank database. In accordance with the phylogenetic results, the present Iranian isolate showed a higher degree of interspecific divergence (up to 3.3%) compared to the data for the taxa available in the GenBank database. Thus, molecular data indicate that this isolate may represent a new species. However, further genetic analyses are needed as a complementary tool to the morphological characterisation in order to elucidate the phylogenetic relationships of Haemogregarina spp.

Coenurosis is a debilitating disease caused by the larval stage of Taenia multiceps. The larval stage of T. multiceps appears in two cerebral and non-cerebral forms known as Coenurus cerebralis and Coenurus gaigeri,respectively. This leads to the hypothesis that there are genetic intraspecific variabilities within this species. Molecular analysis in the present study showed that C. cerebralis and C. gaigeri were 100% genetically identical based on exonic region of enolase (ENO) gene and both mitochondrial (cox1 and nad1) markers. In addition, an intronic mutation at ENO gene (position: 1171) was detected in the Iranian C. cerebralis. The phylogenetic analyses based on the ENO, cox1, nad1 sequence data also suggest that T. gaigeri may not be distinct from T. multiceps and there is only one single valid species within T. multiceps., (Copyright © 2015. Published by Elsevier B.V.)

In this study, molecular characterization of Echinococcus granulosus sample obtained from water buffalo originating from southwest of Iran was performed using comparative sequence analysis of cox1 mitochondrial gene. DNA was extracted from protoscoleces removed from hydatid cyst from the liver of a 2-year-old male buffalo slaughtered in Khuzestan province. Molecular and phylogenetic analyses were conducted based on cox1 mitochondrial gene. We found the presence of a new microvariant of G3 genotype for E. granulosus in Iran which is genetically differentiated from reference G3 sequence (M84663). The difference was a transition mutation of adenine to guanine in position 214 (A214G) resulting in a substitution of the threonine (ACT) by alanine (GCT). These findings extend the knowledge of heterogeneity and distribution of G3 genotype for E. granulosus in world.

Sheep and goats serve as intermediate hosts for the canine tapeworm Taenia multiceps. The cysts produced by the intermediate stage of parasite are usually found in the cerebral hemispheres of small ruminants, and the resulting disease is commonly known as coenurosis. Coenurosis is clinically manifested in the form of various nervous symptoms, depending on the exact location of the cyst. The variety of neurological symptoms contributes to the complexity of clinical diagnosis and reinforces the need for a more specific and acceptable diagnostic approach. We demonstrated here, for the first time, that the T. multiceps DNA is present in the cerebrospinal fluid (CSF) of the infected sheep and goats. In addition, the molecular genetic marker of the mitochondrial DNA was applied phylogenetically to show that our isolates together with other T. multiceps strains comprised a monophyletic group that is a sister to Taenia krabbei. Pairwise comparison between the cox1 sequences of our study and other T. multiceps genotypes existing in the GenBank showed similarity ranging from 98 to 100%. Accordingly, the polymerase chain reaction (PCR) can be used for amplification of DNA of the parasite originated from the CSF and provides a valuable method for accurate identification of coenurosis cases.

This experimental study was conducted to test whether cerebral and non-cerebral forms of Coenurus cerebralis belong to one origin or they are originated from two different tape worms. In the first step of the study, two groups of dogs were orally infected with the protoscoleces of cerebral and non-cerebral cysts and four months after infection, the adult worms were collected and morphologically characterized. Then the obtained eggs from two groups of adult worms were orally inoculated to two groups of goats to trace the predilection sites and also to compare the morphological and molecular characteristics of the larval stage of the parasites. The results showed that, both cerebral and non-cerebral coenuri, produced morphologically similar worms in the experimentally infected dogs. We observed only non-cerebral coenurus cysts in two groups of experimentally infected goats and the morphological characteristics of the cysts of two different groups were also similar. The molecular findings also showed that the cysts of two groups were 100% identical to each other based on the CO1 and nad1 sequences. In conclusion, our morphological and molecular findings reinforce the evidence that Taenia multiceps is the single origin of both cerebral and non-cerebral form of C. cerebralis but in goats, the cysts show more tendency to non-cerebral organs., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: Fasciola hepatica and F. gigantica are the causative agents of fasciolosis in domestic animals and humans. Based on the morphometric criteria, differential diagnosis between them is problematic. In addition, intermediate forms of Fasciola have been found in Iran, which makes the differentiation more difficult. The aim of the present study was to provide molecular evidence for the existence of F. gigantica in Iran using sequencing analysis of ND1 and PCR-RFLP analysis of ITS2 regions and to study the intraspecies variations of F. gigantica based on mitochondrial ND1 gene polymorphism., Methods: Forty Fasciola spp. samples collected from four distinct provinces (Fars, Khuzestan, Gilan, Khorasan Razavi) in Iran were collected for morphological and molecular characterization. In molecular method, PCR-RFLP analysis of ITS2 using pagI restriction enzyme was used as a screening approach for F. gigantica differentiation. Then mitochondrial DNA sequence variations in the ND1 gene were used for phylogenetic analysis., Results: Based on the morphometric criteria and RFLP analysis, 14 parasitic samples were initially identified to be F. gigantica. Phylogenetic results showed that there are at least 10 different genotypes of F. gigantica in Iran, which are different from those existing in the GenBank. Twenty-six points out of 410 base pairs of sequenced ND1 gene in 10 varieties of F. gigantica were diagnosed to be polymorphic. From 26 points of polymorphism, only eight resulted in the post-translational amino acid changes in ND1 gene product structure., Conclusion: Data revealed noticeable genetic diversity (up to 4.63%) between different varieties of F. gigantica in Iran.

Habronema muscae is a spirurid nematode that undergoes developmental stages in the stomach of equids, causing chronic catarrhal gastritis. Despite preceding investigations have developed polymerase chain reaction (PCR)-based assays for molecular diagnosis, we aimed to assess the applicability of cytochrome c oxidase subunit 1 (cox1) sequences to identify the H. muscae infection and to assess the level of intraspecific variations in this parasite obtained from affected horses in Southern Iran. According to the morphological characterizations, two different isolates of H. muscae were identified. Although the majority of the recovered specimens had normal characterizations of H. muscae, a number of parasites showed an abnormal feature as large, asymmetrical, and thick cuticular extensions was observed at their anterior end (head region) in gross and histologic examinations. Unexpectedly, molecular assay disclosed that both morphologically distinct samples were completely identical to each other based on cox1 sequence. Multiple alignment of the cox1 amino acid sequences showed that all polymorphism sites were silent. Also, phylogenetic analysis provided strong support that H. muscae form a sister group to Spirocerca lupi and Thelazia callipaeda.

Five common tetracycline resistance genes tet(A), tet(B), tet(M), tet(O) and tet(S) were studied by polymerase chain reaction in 100 bacteria isolated from Iranian fish farms. In the antibiogram test most of the bacteria were either intermediately or completely resistant to tetracycline. Nine isolates out of 46 Aeromonas spp. contained either tet(A/M/S) resistant genes as follows: tet(A) in A. veronii/sobria (n = 1), A. media (n = 2), A. aquariorum (n = 1), and A. veronii (n = 3); tet(M) in one isolate of A. sobria and tet(S) in 1 isolate of A. jandaei. In other bacteria, tet(A) gene was detected in Citrobacter freundi (n = 1), Pseudomonas putida (n = 1); tet(S) was also identified in Yersinia ruckeri (n = 1), Arthrobacter arilaitensis (n = 1) and P. putida (n = 1). In total, 31 isolates (31.00%) contained the tetracycline resistance genes in which 21 bacteria (21.00%) showed the tet(S), nine bacteria (9.00%) contained the tet(A) and 1 bacteria (1.00%) was positive for tet(M). All of the L. garvieae isolates contained tet(S) in this study. The most widely distributed resistance gene was gene tet(A) and the least known resistance genes was tet(M) among the studied bacteria of the genus Aeromonas in this study.

Background: Campylobacter is one of the leading bacterial species causing foodborne illnesses in humans. Antimicrobial agents have been extensively used for treatment of Campylobacter infections; but in the recent years, both animal and human isolates of this bacterium have shown resistance to several antibiotics such as tetracycline., Objectives: The aim of this study was to investigate the presence of genetic determinants of tetracycline resistance in Campylobacter spp. recovered from poultry carcasses in Shiraz, Iran., Materials and Methods: Eighty-three thermophilic Campylobacter spp. Isolates were first identified based on multiplex polymerase chain reaction (PCR) and then screened for presence of tetracycline resistance genes (tet (A), tet (B), tet (O) and te (S)) by PCR., Results: The overall prevalence of Campylobacter jejuni and C. coli among the examined isolates was 51.8% and 48.2%, respectively. Tetracycline resistance genes of tet (B) and tet (S) were not seen among these Campylobacter spp. Isolates, whereas the most common tet gene identified was tet (O), found in 83.1% (69/83) of all the isolates. The tet (O) gene sequence comparison between C. jejuni and C. coli showed 100% similarity and these sequences (JX853721and JX853722) were also identical to the homologous sequences of other strains of Campylobacter spp. existing in the GenBank databases. In addition, tet (A) was found in 18% (15/83) of Campylobacter spp. isolates. To our knowledge, this represents the first report of tet (A) in Campylobacter spp. There was 100% homology between the sequences of tet (A) from this study (JX891463 and JX891464) and the tet (A) sequences mentioned for other bacteria in the GenBank databases., Conclusions: The high prevalence of tet (O) resistance gene along with new detection of tet (A) resistance gene in Campylobacter spp. isolated from poultry carcasses revealed an extensive tetracycline resistance among Campylobacter isolates from poultry in Iran. It emphasized the need for cautious use of tetracycline in poultry production to decrease the extension of tetracycline-resistant Campylobacter spp.

Hemoplasmas are the trivial name for a group of erythrocyte-parasitizing bacteria of the genus Mycoplasma. This study is the first report of hemoplasma infection in Small Indian Mongoose (Herpestes Javanicus) based on molecular analysis of 16S rDNA. Whole blood samples were collected by sterile methods, from 14 live captured mongooses, in the south of Iran. Candidatus Mycoplasma turicensis (CMt)-like hemoplasma was detected in blood samples from one animal tested. BLAST search and phylogenetic analysis of partial 16S rDNA sequence (933bp) of the hemoplasma from Small Indian mongoose (KJ530704) revealed only 96-97% identity to the previously described CMt followed by 95% and 91% similarity with Mycoplasma coccoides and Mycoplasma haemomuris, respectively. Accordingly, the Iranian mongoose CMt isolate showed a high intra-specific genetic variation compared to all previously reported CMt strains in GenBank. Further molecular studies using multiple phylogenetic markers are required to characterize the exact species of Mongoose-derived hemoplasma., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

A 2-year-old male Pekingese dog was referred to Shiraz University's Veterinary Teaching Hospital for anorexia and depression. The case had no history of surgery. Physical examination revealed no abnormalities except mild depression and fever. Small, coccoid, epicellular bacteria were detected on erythrocytes by microscopic examination of the Giemsa-stained blood smears. Abnormalities noted in the complete blood count included regenerative anemia characterized by a marked reticulocytosis. Examination of the plasma showed visual evidence of slight intra vascular hemolysis. In addition, Howell-Jolly bodies, nucleated RBCs, increased immature neutrophils and thrombocytosis were found in this case. The urine was strongly positive for bilirubin, and the urine sediment had abundant bilirubin crystals. For polymerase chain reaction (PCR) purpose, total DNA was extracted from blood sample collected from dog. PCR was positive and phylogenetic analysis of concatenated data showed our isolate clustered within Candidatus Mycoplasma hematoparvum group. Treatment was performed by oral ciprofloxacin and prednisolone. The clinical signs improved after three days. Two month follow-up showed no recurrence. In conclusion, hemoplasmosis should be considered as a differential diagnosis in dogs with hemolytic process and pyrexia. The PCR evaluation for hemoplasma DNA should be included in the investigation of such cases to enable the rapid detection of this infection, which may be more common than previously estimated. Besides, ciprofloxacin might have an effect on treatment of hemoplasma in dogs, however, conducting further case studies are necessary to recommend successful treatment.

Vertical and consequently horizontal transmission of quinolone and fluoroquinolone resistant Escherichia coli clones following hatch in chickens enables a massive amplification of these clones into a large population. The aim of this study was to determine the antibiotic resistance and susceptibility of Iranian E. coli isolates (n=105) from one-day-old chicks to fluoroquinolones and the relation of this resistance with mutations in gyrA and parC genes using PCR-RFLP. For the first time, EcoRV restriction enzyme was used for rapid mutation screening in parC (Ser80Ile). The results showed that the low level of Minimum Inhibitory Concentration (MIC) for ciprofloxacin (0.25-4μg ml-1) and enrofloxacin (0.25-4μg ml-1) corresponded to a single mutation in gyrA, while intermediary to high level of MIC for ciprofloxacin (8 --> 64 μg ml-1) and enrofloxacin (16 --> 64 μg ml-1) were related to 2 mutations in gyrA or 3 mutations, 2 in gyrA and 1 in parC. There was a strong positive correlation (R = 0.93, P < 0.001) between MIC levels of enrofloxacin and ciprofloxacin among these isolates. The article concludes by stressing that the rising incidence of enrofloxacin resistant E. coli isolates from chicken sources may increase the potential risk of ciprofloxacin resistant E. coli acquisition by humans.