Veterinary Research Forum

Abstract Menthol is an organic compound derived from mint oils, known for its cooling and soothing properties, often used in various pharmaceutical, cosmetic, and therapeutic applications. It exerts beneficial effects on the digestive system by relaxing smooth muscles, reducing spasms, and improving gastrointestinal motility. This study aimed to evaluate the effects of menthol on bovine ileal smooth muscle contractions in vitro. Ileal tissue segments were collected from adult cattle at an abattoir and subsequently immersed in 25 mL organ baths containing Tyrode’s solution. The baths were maintained at 37 °C and continuously aerated with a gas mixture of 95% O₂ and 5% CO₂. The tissues were subjected to various contractile agents, including potassium chloride (KCl) at concentrations of 30 and 80 mM, carbachol (CCh) at 1 μM and 4 μM, and barium chloride (BaCl₂) at 30 mM. Menthol was cumulatively applied in incremental concentrations to assess its modulatory effects on contraction amplitude. Results demonstrated that menthol elicited a dose-dependent inhibition of smooth muscle contractions across most stimulatory conditions, with the extent of inhibition varying among different stimuli. The Ca2+ channel blocking activity was further confirmed when pre-treatment of isolated ileums with menthol (23 and 200 μg mL⁻¹) caused a rightward shift in the Ca2+ concentration-response curves (CRCs), similar to verapamil. These findings suggest menthol’s spasmolytic action may be mediated through the modulation and inhibition of calcium channels. Menthol effectively attenuates bovine ileal smooth muscle contractions in vitro, indicating its potential as a natural therapeutic agent for controlling gastrointestinal hyperactivity in cattle.

Abstract Rotavirus is one of the major causes of diarrhea in different animal species and has a bad economic impact due to the losses in neonates and productivity. To investigate the occurrence of this infection in bovine calves, three localities in Khartoum State, Sudan, were selected. A total of 200 fecal samples were collected from diarrheic calves; 100 from Khartoum and 50 from each of Khartoum Bahari and Omdurman provinces. Collected samples were screened for a group A rotavirus antigen using enzyme-linked immunosorbent assay (ELISA). Positive results were seen in 40.00% of samples; the highest prevalence of 42.00% was found in samples from Khartoum province. Five ELISA-positive samples were examined under electron microscope, and characteristic wheel-like appearance of rotavirus was visualized. Polyacrylamide gel electrophoresis was also applied on 15 of the positive samples; eight samples showed different polyacrylamide gel electrophoretic group A rotavirus long profile with different patterns. The results showed that the occurrence of rotavirus infection in cattle in Khartoum State is increasing.

Abstract Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis, an oncogenic deltaretrovirus that has emerged as a potential zoonotic infection. The BLV naturally infects cattle and causes economic losses through a slow persistent infection with various clinical symtoms following preleukosis. The main objective of this study was to determine the seroprevalence of BLV antibodies in cattle and buffaloes in the border provinces of the Eastern Anatolia region, Türkiye, using the agar gel immunodiffusion (AGID) assay and  enzyme-linked immunosorbent assay (ELISA). For this purpose, a total of 1,033 serum samples were collected from 982 cattle and 51 buffaloes from the provinces of Ağrı (n = 178), Iğdır (n = 252), Kars (n = 317), Van (n = 221), and Hakkari (n = 65) during 2021 - 2022. In AGID and ELISA tests, seropositivity for BLV-specific antibodies was not detected in cattle and buffaloes from the mentioned provinces. This study revealed that BLV was not circulating in cattle and buffaloes in the easternmost border provinces of Türkiye during the sampling period and contributed to determine the status of BLV in the mentioned region. Due to the presence of virus in other regions of Türkiye and neighboring countries, Iran and Iraq, it is recommended to control animal movements, continue efforts to combat the transmission of the virus, and maintain control measures.

Abstract An increase in morbidity during times of relatively high functional load on the body such as during pregnancy, confirms the role of metabolic overload in the occurrence of metabolic failures. It is better to take preventive measures such as adjusting metabolic regulation mechanisms in light of the ideal dietary composition. However, this direction is constrained by the lack of information about neurohumoral regulation. The goal of the present study was to learn more about the dynamics of changes in insulin and glucose levels in pregnant cow’s blood. Research on the levels of glucose and insulin in lactating cows demonstrated that ruminants had distinct characteristics in the metabolism of carbohydrates, most notably a lessened reliance of blood glucose on insulin levels. A rise in blood glucose and insulin levels was observed as the gestational stage increased during the third trimester of pregnancy. The intensity of this insulin tolerance was contingent upon the level of productivity and glucose levels during the preceding stages of gestation.

Abstract The bovine leukemia virus (BLV) is an important infectious agent transmitted from cattle to humans. It is considered one of the oncogenic viruses in breast cancer, so an accurate detection of this virus is important. The study aimed to design a specific and sensitive method based on TaqMan^® real-time polymerase chain reaction (RT-PCR) for BLV detection. Probes and primers were designed using bioinformatics software for a 108 pairs region of the BLV tax gene. Criteria employed for determining analytical sensitivity were prepared using in-vitro RNA transcriptions. The National Center for Biotechnology Information (NCBI), basic local alignment search tool (BLAST) databases various viral panels and genomic samples from healthy individuals (Qom Province, Iran in 2023) were used to verify analytical specificity and clinical specificity, respectively. This method can measure a minimum of 10 copies of DNA and RNA mL^-1. Moreover, the assay is linear in the range of 10⁰ - 10⁹ copies mL^-1. By testing negative specimens, the method specificity was 100%. The reproducibility results of the reaction were examined at the intra- and inter-assay comparison. In fact, 10 technical replicates of each concentration of the control sample were analyzed in each working reaction. Due to the locally made kit, exact sensitivity and specificity, rapid analysis, and relatively low cost, as compared to commercial kits of other countries, the method introduced in the present study could be suitable for accurate detection of the BLV. Also, the TaqMan^® real-time PCR method could be detected in cattle and human and before malignant changes of breast cancer which could reduce infection and breast cancer.

Abstract In November 2021, an investigation was conducted into an outbreak of abortion, stillbirth, and the birth of calves with congenital abnormalities (arthrogryposis and hydranencephaly) at a dairy farm in Dasht-e-Mughan city, Ardabil province. A total of 70 cows experienced these issues. To determine the cause of the outbreak, post-mortem brain tissue samples were collected from two calves affected by hydranencephaly, which occurred shortly after their birth. Polymerase chain reaction (PCR) testing was conducted for multiple viruses, including bovine viral diarrhea (BVD), border disease, Akabane, Schmallenberg, and bluetongue viruses (BTVs). The samples were positive only for Akabane virus. Serum samples were collected from a group of 60 cattle, consisting of 45 adult cows and 15 younger calves aged between 8 to 10 months. These samples were analyzed to detect the presence of antibodies against the Akabane and Schmallenberg viruses. Both of these viruses are known to be responsible for causing abortion, stillbirth, and congenital abnormalities in calves. Among 45 cows that tested by competitive enzyme-linked immunosorbent assay (cELISA), 26.66% and 33.33% exhibited antibodies against Akabane and Schmallenberg viruses, respectively. Notably, 20.00% of cows co-exhibited antibodies for both viruses. Despite PCR evidence implicating Akabane virus as the principal etiology of clinical signs observed in the affected herd, the high co-seropositivity to Schmallenberg virus, warrants a thorough investigation into potential viral interactions. Further research is required to determine the source of the virus and their transmission routes. This information could facilitate the refinement of disease control strategies and improving the management of reproductive challenges in such affected herds.

Abstract The interval between parturition and subsequent pregnancy is called the days open or calving to conception interval and is affected by several factors, especially dystocia. Dystocia is an ab-normal or difficult calving that may require assistance during labor. This study is a field trial in health education and the research team developed a comprehensive training program for farmers to educate them about the normal process of parturition in dairy cows and when and how to assist in parturition or dystocia. A series of classes was held for farmers and the study covered 486 multi-parous dairy cows, with 173 belonging to the group of trained farms (educated farmers) and 313 to the control group (non-educated farmers). Although dystocia was lower in the educated group, there were no significant differences in retained placenta between two groups. However, cows in the educated group had a better conception rate (lower service per conception) in sub-sequent parturitions. Hence, the median number of days open for cows from trained farmers was significantly lower than other farmers (85 days compared to 120 days, respectively). Based on Cox regression analysis, uterine prolapse, retained placenta, and dystocia could significantly impact subsequent pregnancies. Dystocia affects days open, and training on parturition and dystocia management can effectively reduce the numbers of days open in dairy cows.

Abstract The present research was carried out to assess the serum progesterone (P₄) concentration and uterine hemodynamics at estrus till ovulation in cyclic cows (N = 130) with healthy or diseased uterus. At estrus, 85 cows were diagnosed with clinical endometritis (CE; n = 44) and sub-clinical endometritis (SCE; n = 41); whereas, 45 cows being served as control namely no endometritis (NE; n = 45) were included in the study. Serum progesterone estimation at 12 - 14 and 40 hr after the onset of estrus and Doppler sonography of both middle uterine arteries were done to envisage the uterine hemodynamics and ovulation. The serum progesterone concentration was significantly higher at 12 - 14 hr after onset of estrus in CE and SCE cows. At 12 - 14 hr after onset of estrus, a cut-off value of ≥ 0.48 ng mL^-1 P₄ was obtained, above which 22.72% CE, 26.82% SCE and only 8.88% NE cows failed to ovulate within 36 - 40 hr of estrus onset. Among the Doppler indices, pulsatility and resistance indices were significantly higher; whereas, volume and velocity indices were significantly lower in NE cows. In cows diagnosed with CE and SCE, a higher supra-basal P₄ concentration, and velocity and volume of blood flow to uterus at estrus negatively affected the duration to ovulation.

Abstract Tropical or Mediterranean theileriosis in dairy cattle is widely distributed in many tropical regions of the world. The purpose of this study was to evaluate the proliferation status of mononuclear cells infected with Theileria annulata schizonts in different tissues and its relationship with the pathogenesis of the parasite in cattle by histopathology, immuno-histochemistry and polymerase chain reaction (PCR). Blood and tissue samples of eight Holstein cattle that had been lost due to theileriosis and eight healthy slaughtered cattle of the same breed were collected as a control group after necropsy. The piroplasms in the blood smears and the schizonts in the cytoplasm of the lymphocytes and macrophages of the lymph nodes were microscopically detected. Histopathologically, the proliferation of macrophages, lymphocytes, and plasma cells in lymph nodes and the heart, congestion, and bleeding in the red pulp of the spleen, portal tracts of the liver, interstitial tissue of the kidneys, multifocal necrosis and ulceration in the abomasum together with hyperemia and hemorrhages and lymphoblastic infiltration in the submucosa and lamina propria adjacent to these lesions and emphysema with ecchymotic hemorrhage in the lungs were evident. Immunohistochemistry identified the proliferated cells as mostly Cluster of Differentiation 3- Positive T lymphocytes and macrophage marker antibody 387- positive macrophages. Positive results of PCR for the Tams1 30.00 kDa gene were observed in lymph nodes, liver, lung and abomasum. It was concluded that the pathological changes were the result of schizont-infected macrophage proliferation leading to severe uncontrolled proliferation of uninfected T lymphocytes.

Abstract Foot-and-mouth disease (FMD), a highly contagious viral disease of livestock, is endemic in Iran. To investigate the prevalence of antibodies against 3ABC non-structural protein (NSP) of FMD virus, a cross-sectional study was conducted on dairy cattle in eight cities of Kurdistan Province from May to September 2016. Serum samples (n = 283), were collected from cattle vaccinated with the recommended dose of a commercial vaccine and tested by a Competition enzyme-linked immunosorbent assay. Results showed the overall seroprevalence of antibodies against NSP of FMD virus in the vaccinated cattle was 22.30% (95.00% CI: 17.40 - 27.20%). The seroprevalence of antibodies was affected by geographical regions, with the highest seroprevalence related to the samples of vaccinated cattle in the cities of Marivan 95.00% (95.00% CI: 92.50 - 97.50%) and Saqqez 38.50% (95.00% CI: 32.80 - 44.20%). In terms of age, the highest seroprevalence of antibodies to FMD virus 26.70% (95.00% CI: 21.60-31.80%) belonged to ≤ 24-month-old cattle. These findings suggest that the presence of NSP antibodies in vaccinated cattle indicates the risk of infection with FMD virus serotypes circulating in the west of the province, so further studies with a larger sample size are recommended.

Abstract This study aimed to investigate the potential presence of bovine herpes virus type 1 (BHV-1) and bovine viral diarrhea virus (BVDV) in cattle uteri that did not display any clinical and macroscopic signs of infection. Virus detection involved polymerase chain reaction (PCR) test, double immunohistochemistry (IHC), and double immunofluorescence (IF). One hundred cornu uterus samples were collected from cattle aged 1 year and older. The BVDV was detected by PCR or by double IHC/IF in the collected samples from slaughterhouses in Kayseri city (Central Anatolia, Türkiye) from 2021 - 2022. By contrast, BHV-1 was detected by PCR and double IHC/IF at a rate of 16.00% and 21.00%, respectively. In the IHC and IF detection, BHV-1 was detected in endometrial epithelial cells and in some mononuclear cells in the lamina propria, periglandular areas and myometrium. Although no macroscopic lesion was found in the BHV-1-positive samples (n = 21), histopathological detection showed that two had acute endometritis, eight had subacute endometritis, eight had chronic endometritis and the three others showed no signs of endometritis. This prevalence study demonstrated for the first time that even while BVDV could not be detected in the samples, BHV-1 posed a critical potential reproductive risk in pregnant animals, as it can specifically cause abortions when it resides in cattle uteri that do not show clinical or macroscopic and even microscopic signs of infection. Additionally, this study was the first to combine PCR and double IHC/IF for BHV-1 and BVDV detection in cattle uteri.

Abstract The purpose of this study was to investigate the tetracycline resistance in Trueperella pyogenes isolates from bovine samples in Burdur, Turkiye, and assess 16 tetracycline-resistance genes distribution among the isolates. Forty-nine T. pyogenes isolates were phenotypically characterized for anti-microbial resistance to doxycycline, oxytetracycline and tetracycline by disc diffusion method. Presence of tetracycline genes of T. pyogenes was investigated by multiplex and singleplex polymerase chain reaction. Our results indicated that 87.80% and 42.86% of the isolates were resistant to tetracycline and oxytetracycline, respectively, and the rate of resistance to doxycycline was 6.12%. Total of 21 (42.85%) were carrying tetracycline-resistance genes and tet(A) was present in 12 (24.49%) isolates; whereas, the tet(W) gene was identified in 9 (18.37%) and 2 (4.08%) of the isolates carried both tet(A) and tet(W), respectively. The study indicated antibiotic resistance patterns of tetracycline agents and links to the tet-genes among T. pyogenes were detected. It makes it worthwhile that this is the first report for detection of tet(A) gene in T. pyogenes.

Abstract Despite being important, there are no equations for prediction of ionized calcium (iCa) in sheep and cattle. The objectives of this study were i) to create equations for the calculation of serum iCa concentration based on the serum concentrations of total calcium (tCa), albumin (Alb) and total proteins (TP) and ii) to investigate whether predicted serum iCa values are beneficial in clinical practice. Serum samples from 30 sheep and 30 dairy cattle were used. Serum tCa was determined colorimetrically, while serum iCa was determined with an ion selective electrode method. Serum Alb and TP concentration were determined using bromo-cresol green and biuret methods, respectively. Ionized calcium was also calculated based on serum tCa, using regression analysis, and with two equations based on Alb and TP concentration. Bland–Altman plots were plotted to evaluate the agreement between measured and predicted iCa; Passing and Bablok (P - B) regression analysis was used to assess their agreement. The initial equations were corrected using the P - B generated equation and Bland – Altman plots were run to evaluate the level of agreement between measured and predicted iCa using the final equations. Six equations were finally created for cattle and 6 for sheep. The total bias exceeded 10.00% in all of them indicating that they are clinically unacceptable for iCa prediction especially when the predicted result is very close to the cut-off point of < 1.00 mmol L^-1. So, it could be suggested that, when necessary, iCa concentration should be directly determined.

Abstract Toxoplasmosis, a foodborn disease, in human occurs commenly after the ingestion of tissue cysts via the raw and/or undercooked meat of different infected intermediate hosts such as sheep and cattle.The current study aimed to detect the genetic structure of Toxoplasma gondii isolated from various organs of sheep and cattle in the north of Iran. Convetional PCR was carried out by B1 and REP-529 genes of T. gondii. Nested and RFLP-PCR were performed for all positive samples using SAG2 and GRA6 genetic markers. Amplicons from second round of nested-PCR were sequenced and analyzed with NCBI database. Among of 179 examined samples, 38(21.20%) were positive. The highest of positive cases were found in kidney (28.60%). PCR-RFLP of SAG2 and GRA6 genes demonstrated the alleles of clonal type III in the all of isolates. Sequence analysis of the amplicons revealed the alleles of clonal type III and atypical isolates (Tg-67, Tg-100 and Tg-106). Phylogenetic analyses showed separate clade for the atypical isolates from others in the present study and the reference strains clades. In conclusion, the genetic characterization of T. gondii isolates from sheep and cattle showed high genetic diversity compared with standard type I, II and III genotypes. These results support the hypothesis of the existence of polymorphic and overlapping strains within livestock in Iran. It also suggested the necessity of increased genotyping and sampling efforts to accurately estimate T. gondii intra specific genetic diversity

Abstract Schmallenberg virus (SBV) is an emerging single-stranded RNA virus being classified under Simbu serogroup of Bunyaviridae family. This study aimed to detect antibodies against SBV in cattle for the first time in three eastern provinces of Iran. Blood samples were randomly collected from jugular veins of 270 cattle, from 19 farms in Razavi Khorasan, South Khorasan and Sistan and Baluchistan provinces. Separated sera were analyzed to find SBV antibody using ID vet^® SBV indirect multi-species enzyme-linked immunosorbent assay test kit. From a total of 273 serum samples analyzed for SBV presence, 12.45% (n = 34) were positive for SBV antibody. Risk factors including breed, age and geographic area showed a statistically significant relationship with the virus prevalence. In conclusion, the seroprevalence of SBV is not high; but it is considerable in the studied parts of Iran. This is the first study regarding SBV seroprevalence in cattle population of eastern Iran and further studies about the virus epidemiology are recommended.

Abstract The main objectives of this study were to determine the occurrence and potential causative factors of Immune-mediated hemolytic anemia (IMHA) in native cattle and water buffaloes from southwest of Iran. Fifty-three anemic animals (37 cattle and 16 buffaloes) were studied. A full clinical history and physical examinations were undertaken for all animals. Four clinically healthy cattle and four healthy buffaloes were also used as control animals. Blood samples were subjected to a complete blood count, Coombs’ test, erythrocyte osmotic fragility test and serum biochemical analysis. IMHA was diagnosed in 12 (32.43%) cattle and 6 (37.50%) buffaloes based on the Coombs’ test. Underlying or concurrent diseases, including theileriosis, anaplasmosis, vaccination, and pneumonia were detected in 11 cattle and four buffaloes. Primary or idiopathic IMHA was identified in one cattle and two buffaloes that their Coombs’ test was positive. Hematologic and biochemical findings in the cattle with IMHA included a nonregenerative anemia, leukopenia, thrombocytopenia, increased osmotic fragility, hyperbilirubinemia and elevated serum alkaline phosphatase, aspartate aminotransferase and lactate dehydrogenase activities. It can be concluded that IMHA occurs in a significant proportion of anemic cattle and river buffaloes in southwest of Iran. The occurrence of IMHA in both cattle and buffaloes is mostly secondary to infectious diseases especially theileriosis and anaplasmosis. Clarification of the mechanisms of primary or idiopathic and secondary IMHA in cattle and buffaloes require further studies.

Abstract Giardia duodenalis is one of the most prevalent intestinal protozoa infecting humans and domestic animals. The aim of this study was to identify subspecies of G. duodenalis by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method from fecal samples of naturally infected cattle in the Urmia, West Azerbaijan province, Iran. Overall, 246 fecal specimens were collected from the cattle (diarrheic and healthy) and microscopically examined for G. duodenalis. The PCR-RFLP analysis of glutamate dehydrogenase (gdh) locus was used to identify the genotypes found in cattle. In this method, 432 bp expected size was amplified and then specific restriction NlaIV enzyme was used for subspecies detection. Totally, 23 (9.34%) specimens were microscopically positive for giardiacyst out of 246 examined samples. The PCR-RFLP analysis revealed that 19 samples (82.60%) have the genotype E and 4 samples (17.39%) belong to the subgroup AI. Our findings indicated that G. duodenalis infection is prevalent in cattle of Urmia and the non-zoonotic genotype E predominates in cattle in this region.

Abstract A total number of 450 blood samples were collected from 45 different randomly selected cattle herds. Light microscopic examination of blood smears revealed Babesia spp. infection in 4.2%, while 8.9% of blood samples were positive using PCR. Upon multiplex-PCR (mPCR), B. bigemina and B. bovis infections were detected in 37/40 (92.5%) and 3/40 (7.5%) samples, respectively. 530 ticks of 10 Ixodid species were collected from the same cattle. Hyalomma anatolicum was the most prevalent tick species (19.9%). An expected 520 bp fragment of Babesia spp. was generated in 22 (48.8%) of Rhpicephalus annulatus, 18 (40.0%) of R. bursa and 12 (30.0%) R. sanguineus sensu lato. The mPCR findings revealed that all infected ticks including R. annulatus, R. bursa and R. sanguineus were totally infected with B. bigemina. The DNA amplification of B. bovis and B. bigemina in egg samples showed that only B. bigemina was detected in two specimens of R. annulatus. It could be concluded that B. bigemina was the dominant causative agent in this region but the evidence of B. bovis infection of cattle in a few cases was noted, as well. The results suggested that B. bigemina and B. bovis could be detected in the DNA extracted from R. annulatus, R. bursa and R. sanguineus sensu lato confirming previous reports. Since B. bigemina is transmitted transovarially by R. annulatus, it might act as an important vector for B. bigemina.

Abstract Members of gram-negative bacteria family Pasteurellaceae, include a large number of important economically human and veterinary pathogens. Organisms belonging to the family can colonize in mucosal surfaces of the respiratory, alimentary, genital tracts and cause diseases in various mammals, birds, and reptiles. Hemorrhagic septicemia is an acute disease of cattle and buffaloes in tropical countries caused by Pasteurella multocida serotype B:2. In the present study, the possible bactericidal activity of immune calf sera in the presence and absence of complement system was investigated. The results showed that P. multocida B:2 is highly resistant to positive serum, containing high levels of IgG and IgM obtained from calves after vaccination, and complement activity in normal fresh calf serum. This organism also grew rapidly in the normal fresh calf serum and the mixture of positive serum as well as normal fresh calf serum. As a control test an E. coli strain was subjected to the same experiment and found completely sensitive to the bactericidal activity of complement in calf and guinea pig fresh sera. Results were indicative of the presence of inhibitory mechanism(s) in P. multocida B:2 against bactericidal activity of immune calf serum and complement system.

Abstract Althoughpoultry meat is considered as the main source for human Campylobacter infections,there is limited information about non-poultry sources. The present study was aimed to investigate the prevalence and the antibiotic resistance of thermophilic Campylobacter spp. in fecal samples of the cattle and sheep in Shiraz, Iran. A total of 302fecal samples were obtained from clinically healthy, slaughtered cattle and sheep from Shiraz slaughterhouse. The animals were clinically healthy before being slaughtered. The samples were cultured according to the specific cultivation method under thermophilic conditions. The susceptibility of Campylobacter isolates were determined for 13 antimicrobial agents. All enriched samples and cultured isolates were targeted for polymerase chain reaction (PCR) detection of 16S rRNA and multiplex PCR for determining their species. Among 302 fecal samples, 65 (21.5%) and 205 (67.8%) samples were positive for the presence of Campylobacter species with the cultivation and PCR techniques, respectively. All 65 distinct isolates were susceptible to neomycin and colistin and the isolates showed high resistance to cephalotin (83.0%) and ciprofloxacin (67.7%). After the multiplex PCR, 78.5% of total positive samples showed the simultaneous presence of Campylobacter jejuni and Campylobacter coli. In conclusion, the results emphasized that non-poultry farms are important as a possible source of Campylobacter infections.

Abstract This study was carried out to determine the presence and frequency of Anaplasma ovis and Anaplasma marginale in sheep and dairy cattle in West-Azerbaijan province, Iran. A total number of 200 blood samples were randomly collected via the jugular vein from apparently healthy cattle (100) and sheep (100). The extracted DNA from blood cells was screened using genus-specific (Anaplasma spp.) nested-polymerase chain reaction (PCR) based on 16S rRNA gene primer sets. Species-specific PCR was set up using major surface protein 4 (MSP4) gene primer set. None of cattle blood samples were positive for Anaplasma spp. by the first nested PCR. Five samples among the 100 sheep blood samples were both positive in the first nested PCR and A. ovis -specific PCR, based on MSP4 gene. In total, 5.00% of animals were A. ovis positive. This study identified a low prevalence of A. ovis in the blood of apparently healthy sheep in West Azerbaijan province.

Abstract The objective of the present study was to determine the grades of nonspecific bacterial infection of genitalia of repeat breeding cattle by a simple and rapid test under field condition. For this purpose, a total of 100 crossbred Jersey cows comprising of 80 repeat breeding animals presented for treatment and 20 normal cyclic (control group) animals presented for artificial insemination at their first service were selected. Estrual cervical mucus from all the animals was collected at 8 to 12 hr after the onset of behavioral estrus and subjected to white side test (WST) and bacteriological examination. The results of WST showed only 15% of control group had infection but the remaining 85% were free of it. In contrast, the majority of repeat breeding animals (57/80) showed infection (71.25%) and only 28.75% animals were free of infection. In bacterial culture, 60 (75.00%) from the 80 repeat breeding animals were found positive, and 20 (25.00%) were free of bacteria. All the three samples of control group that showed no color reaction in WST had also no growth in bacterial culture. The WST results showed a positive (p < 0.01) correlation of 0.48 with bacterial culture. It is thus concluded that under field condition WST can be used as a prime modality for ascertaining nonspecific bacterial infection of repeat breeding cattle before subjecting them to any antibiotic therapy thereby reducing the cost of diagnosis and treatment.

Abstract Fasciolosis is a disease caused by liver fluck of the genus of fasciola. Diagnosis of fasciolosis has been challenging for a long period due to low sensitivity of the coprological diagnostic method. In this study, an in-house Dot-ELISA method; using excretion–secretory (ES Ag) and Crude (Cr Ag) antigens of fasciola was described for diagnosis of fasciolosis in cattle. For this purpose, the sera specimens of slaughtered cattle were taken and examined for fasciola infection. Sera from two groups of cattle, one infected with fasciola (n = 60) and the other non-infected with fasciola (n = 60), were used in the Dot- ELISA test. All sera were tested and evaluated. Except specificity, other parameters such as, sensitivity, accuracy, positive and negative predictive values of Dot- ELISA with ES Ag were better than those of Dot- ELISA with Cr Ag. In conclusion, excretory–secretory antigen dependent Dot-ELISA can be used as a reliable sero-diagnostic test for fasciola infection in cattle.

Abstract Bovine neosporosis caused by the apicomplexan protozoan parasite N. caninum, was initially recognized in 1989 and is now reported as a leading infectious cause of reproductive failure in dairy cattle in world wide. The aim of this study was to determine the seroprevalence of N. caninum infection in industrial dairy cattle of Hamedan province (west of Iran) by ELISA method. Blood samples were collected from 492 cattle in 41 farms. Antibodies to N. caninum were found in 63(12.80%) sera. A Significant difference was observed between seropositive cattle and dog presence in farm, dog contact with herd, abortion history and herd population. No significant differences were found between seropositive cattle and age as well as breed. This study is the first report of N. caninum infection in dairy cattle farms in Hamedan province. As per our knowledge, Neospora is an important factor in abortion of cattle in this region. Therefore, comprehensive studies for control strategies and improving management of dairy farms is necessary.

Abstract Electrocardiography (ECG) may be used to recognize cardiac disorders. Levels of milk production may change the serum electrolytes which its imbalance has a role in cardiac arrhythmia. Fifty high yielding and fifty low yielding Holstein dairy cows were used in this study. Electrocardiography was recorded by base-apex lead and blood samples were collected from jugular vein for measurement of serum elements such as sodium, potassium, calcium, phosphorous, iron and magnesium. Cardiac dysrhythmias were detected more frequent in low yielding Holstein cows (62.00%) compared to high yielding Holstein cows (46.00%). The cardiac dysrhythmias that were observed in low yielding Holstein cows included sinus arrhythmia (34.70%), wandering pacemaker (22.45 %), bradycardia (18.37%), tachycardia (10.20%), atrial premature beat (2.04%), sinoatrial block (2.04%), atrial fibrillation (8.16%) and atrial tachycardia (2.04%). The cardiac dysrhythmias were observed in high yielding Holstein cows including, sinus arrhythmia (86.95%) and wandering pacemaker (13.05%). Also, notched P wave was observed to be 30% and 14% in high- and low- yielding Holstein cows respectively. The serum calcium concentration of low yielding Holstein cows was significantly lower than that of high yielding Holstein cows. There was not any detectable significant difference in other serum elements between high- and low- yielding Holstein cows. Based on the result of present study, could be concluded that low serum concentration of calcium results to more frequent dysrhythmias in low yielding Holstein cows.