BACKGROUND: Bovine Herpes Virus-1 (BHV-1) belongs to the  Alpha herpesviral family. The virus is the cause of Infectious Bovine Rhinotracheitis (IBR) and Bovine Abortion. In the initial infection, the virus proliferates excessively. Moreover, shedding the virus leads to conditions in the latent phase of the disease. Infectious Bovine Vulvovaginit (IPV ) is the genital form of the disease that represents a genital infection and transmits via pustules and mucopurulent secretions. Exposure to the virus in genital mucosa leads to IPV infection through mating or artificial insemination and the diseases that can be transmitted to healthy livestock by frozen sperm during artificial insemination.OBJECTIVES: Viral contamination of the semen is one of the routes to spread the disease among dairy cattle. Therefore, we investigated the presence of the virus in domestic and frozen imported semen consumed in industrial dairy cattle farms.METHODS: In the present study, 140 frozen straws were collected. After melting each straw, 200 µl of obtained semen was used for DNA extraction, which was done directly on the semen samples and via a Genome Extraction Kit. Subsequently, to ensure the accuracy of the extraction, the PCR technique was done using PRM-1 gene primer. Tracking the viral genome was done using the PCR technique and known primers.RESULTS: In total, one out of 140 samples was found to be virally contaminated, and IBR contamination was confirmed by repeating all the steps and determining the gene sequence.CONCLUSIONS: It is necessary to further investigate the possibility that contamination can be transmitted via frozen semen, given that even one out of 140 samples is contaminated, and the importance of the disease.

BACKGROUND: Diarrhea is a common disease in the neonate calf which imposes significant economic burden on cattle industry around the world. During the first week after birth, Enterotoxigenic E.coli (ETEC) strains carrying F5 fimbria are one of the most important pathogens causing calf diarrhea. F5 fimbria is involved in early stage of pathogenesis and is responsible for attachment of bacteria to enterocytes; this attachment is mediated by FanC protein of F5 fimbria. Antibodies directed against F5 fimbriae play a significant role in prevention and control of the disease. Objectives: Evaluation and expression of recombinant expression of F5 Fimbriae of Enterotoxigenic Escherichia Coli associated with calf diarrhea. Methods: In the present study,  the fanC region of F5 fimbria was cloned in a pET28a plasmid. Results: The recombinant construct was confirmed by sequencing and protein production in Escherichia coli BL21 (DE3) was evaluated by western blotting procedure. Conclusions: Based on our findings, the recombinant FanC protein or the BL21 (DE3) strain are suitable candidates to develop an effective vaccine against calf colibacillosis or use in a diagnostic kit for F5+ ETEC.

BACKGROUND: Brucellosis is one of the most common zoonosis in Middle East and Iran. OBJECTIVES: The purpose of this study was genomic detection of Brucella spp. in sero-positive dairy cattle. METHODS: We have collected 28,519 blood samples from cows during 2012-2013. Samples were screened by Slide and tube agglutination and 2-Mercaptoethanol tests. Samples with anti-Brucella antibodies titer ≥ 1:80 and ≥1:40 in tube agglutination and 2-ME tests were considered as positive respectively. Tissue samples include: lymph nodes, liver, testicle and kidney from 122 samples of slaughtered cows were collected. The Sero-positive samples were examined by a collection of specific primers for Brucella abortus, Brucella melitensis, vaccinal strains included RB51 and Rev1 using PCR tests. RESULTS: Results showed that 450 samples were positive in slide agglutination test and 447 samples had anti-brucella antibodies titer equal to or more than 1:80. So they were positive by tube agglutination test. Three hundred eighty nine samples were positive by 2- mercaptoethanol test. PCR test results showed that 46 samples (37.7%) out of 122 samples had a specific sequence of Brucella or otherwise they have an active infection with Brucella species, whereas 62.3% of samples were negative. The PCR results showed that 2 samples (4.35%) were infected by B. melitensis, 2 samples (4.35%) infected by Rev1 strain and 42 samples (91.3%) were infected by B. abortus. CONCLUSIONS: The results showed that, as we had expected, the majority of cows were infected by B. abortus. Animals who infected by B. melitensis and Rev1 strain may be a result of contact with sheep or goats. We couldn’t find Brucella genome in 76 samples (62.3%) of sero-positive cows. It may be caused by cross reaction of sera with Brucella species in tests or activation of immune system response and elimination of organism from internal organs.

The present study describes the clinical, serological and bacteriolological findingsin calves from two beef herds experiencing outbreaks of pneumonia. The clinical signs were nasal discharge, cough, pyrexia and increased respiratory rates. The morbidity and mortality rates over a month period were 40.72% and 15.63% respectively. Laboratory investigations revealed that bovine viral diarrhea virus (BVDV) was involved in and probably initiated both outbreaks as indicated by a significant increase in antibody titers against BVDV in sera of convalescent calves (paired serum samples). No antibodies bovine herpesvirus-1 (BHV-1), bovine respiratory syncytial virus (BRSV) and parainfluenza-3 (BPIV-3) viruses were detected in both acute and convalescent sera. Mycoplasma bovis was concurrently demonstrated in lungs of affected calves as it was isolated from 13 (81.25%) of examined lungs suggesting that there may be a synergism between bovine viral diarrhea virus and Mycoplasma bovis in the pathogenesis of pneumonia. A total of 15 (68.18%) isolates of Mannheimia haemolytica, 5 (22.73%) Pasteurella multocida, 1 (4.54%) Pseudomonase aerugenosa, 3 (13.64%) Staphylococcus aureus, 3 (13.64%) Actinomycis pyogenes, 1 (4.54%) Klebsiella pneumonae, 1 (4.54%) Streptococcus pneumonae, 2 (9.09%) E. coli and 2 (9.09%) Aspergellus fumigatus were recovered from lungs of calves suffering from pneumonia.

Lymph nodes (Ln) are the preferred samples for virus isolation and detection. Inthe present study, carcass and visceral Ln of apparently healthy cattle were screened for the presence of bovine herpes virus type 1 (BHV-1) and bovine ephemeral fever virus (BEFV) antigens. A total of 198 Ln (114 carcasses Ln and 84 visceral Ln) were collected. Lymph node homogenates were assayed by agar gel precipitation test (AGPT), rapid Staphyloccocal protein A (SPA) agglutination test and Dot-ELISA. The overall results revealed that BHV-1 antigens were detected in 43.9%, 56.1% and in 68.4% of carcass Ln, and in 29.8%, 47.6% and 57.1% of visceral Ln collected from slaughtered cattle by AGPT, SPA agglutination test and Dot-ELISA respectively. On the other hand, BEFV antigens were detected in 5.3%, 38.6% and 52.6% of carcass Ln, and in 6%, 41.7% and 54.8 % of visceral Ln collected from slaughtered animals by AGPT, rapid SPA agglutination test and Dot-ELISA respectively. The results showed high percentage of positive samples with SPA agglutination test and Dot- ELISA in comparison to AGPT for both BHV-1 and BEF.

Abortions in domestic ruminants cause significant economic losses to farmers. Determining the cause of an abortion is important for control efforts, but it can be challenging. All available diagnostic methods in the bacteriology laboratory should be employed in every case due to the many limiting factors (autolysis, lack of history, range of samples) that complicate the investigation process. The purpose of this study was to determine whether the recovery of diagnostically significant isolates from domestic ruminant abortion cases could be increased through the use of a combination of the existing aerobic culture and Brucella selective method with methods that are commonly recommended in the literature reporting abortion investigations. These methods are examination of wet preparations and impression smears stained by the modified Ziehl–Neelsen method, anaerobic, microaerophilic, Leptospira, Mycoplasma and fungal culture. Samples of placenta and aborted foetuses from 135 routine clinical abortion cases of cattle (n = 88), sheep (n = 25) and goats (n = 22) were analysed by the new combination of methods. In 46 cases, bacteria were identified as aetiological agents and in one case a fungus. Isolation of Brucella species increased to 7.4% over two years compared with the previous 10 years (7.3%), as well as Campylobacter jejuni (n = 2) and Rhizopus species (n = 1). Salmonella species (5.9%) and Trueperella pyogenes (4.4%) were also isolated more often. In conclusion, the approach was effective in removing test selection bias in the bacteriology laboratory. The importance of performing an in-depth study on the products of abortion by means of an extensive, combination of conventional culture methods was emphasised by increased isolation of Brucella abortus and isolation of C. jejuni. The combination of methods that yielded the most clinically relevant isolates was aerobic, microaerophilic, Brucella and fungal cultures.

Xenografting using bovine jugular vein (BJV) (Vv.jugulars) valved conduit is recently introduced for reconstruction of different cardiovascular disorders. Commercially prepared conduit needs a standard technique for selection of suitable animal and jugular vein segment with ideal characters of Valved conduit. The present study was carried out on 10 adult healthy animals (5 cattle (Bos-Taurus) and 5 buffaloes (Bubalus bubalis)) and 10 cadaveric BJV specimens collected from slaughtered healthy adult animals (5 cattle and 5 buffaloes). The study aimed to establish a standard method for choosing the suitable animal and segment of JV that would be used for post-slaughtering collection of conduits. Ultrasonographically, morphological and histological characteristics of JV in cattle and buffaloes were also studied and compared. Ultrasonography of JV was performed along its length from the mandible till the thoracic inlet. The assessed ultrasonographic JV features included; vein lumen width (VLW), JV wall thickness (VWT), distance between valves and venous wall gray scale analysis. Ultrasonographically, venous tricuspid valve appeared in both planes (sagittal and transverse) of the 4th quarter segment in both animals. The VLW; significantly increase in cattle than buffaloes. The VWT; significantly increased in buffaloes than in cattle. Morphologically, CJV has less thick and less tough wall; and wider lumen when compared with BJV. Histologically, JV wall is 3-layered; tunica intima (inside), tunica media and tunica adventitia (outside) and the wall thickness of BJV are thicker than CJV. In conclusion, the 4th quarter of CJV is the most appropriate segment advised for post-mortem collection of JV conduits. Ultrasonography is an essential, prerequisite technique for choosing the suitable animal and the perfect segment of JV conduits.

A new and effective protocol to culture bovine embryos without coculture and with individualized culture media has been established, which would allow the study of a single embryo’s metabolism. For this purpose, bovine embryos were produced in vitro by standard protocols in three different types of media: KSOM, SOFaa, and KSOM followed by SOFaa at day 2. Presumptive zygotes were divided into six groups: control, cultured in groups (C-KSOM, C-SOFaa, and C-KS), and individual well system (W-KSOM, W-SOFaa, and W-KS). Cleavage and blastocyst rates were assessed on days 2 and 7 respectively. Relative quantification of transcripts related to important metabolic processes (GLUT1, GLUT3, GSK3, SOD1, HSPD1, G6PD) were assessed in C-KS and W-KS blastocysts. Results show that cleavage was significantly higher only in W-KSOM when compared to C-KSOM, while blastocyst rates differ only between C-SOF and W-SOF. All the other comparisons did not present statistical difference. Moreover, gene expression analysis revealed that blastocysts cultured in groups and in the individual well system present similar transcription patterns. Thus, the obtained conclusion was that the individual well system performed could be used as an effective alternative protocol for individual culture of bovine embryos, since the rates are similar to routine group culture.

A disease characterized by papules, nodules, vesicles, pustules and ulcers on teats and udder as well as drastic drop in milk production was seen among a cattle farm in Fayoum Governorate, Egypt. A virus was isolated by inoculation of vesicle and scrap homogenate pool from infected cattle into the chorioallantoic membrane of specific pathogen free embryonated chicken eggs. The virus was identified by presence of pock lesions, intracytoplasmic inclusion bodies on the chorioallantoic membrane, polymerase chain reaction and immunohistochemistry of the inoculated membrane. A novel pathogenicity model was developed via ear pinna inoculation of Swiss mice. The virus produced vesicular and ulcerative lesions at the site of inoculation in inoculated mice. The virus identity was confirmed by the presence of intracytoplasmic viral antigens by immunohistochemistry

Forty-eight pathogen free New Zealand rabbits were divided into two groups, the first group contained eighteen rabbits served as normal control and the second group of thirty rabbits were received 1 ml bovine herpesvirus-1 (BHV-1) virus suspension (107 TCID 50) by intraperitoneal route. Rabbits both groups were subjected to hematological, serum biochemical, different serological and histopathological examination 3,7,10,14,21 and 28 days post infection. Clinical observation of infected rabbits showed febrile response and mild conjunctivitis after 24 and 48h. of inoculation, respectively. The hemogram revealed no significant alteration in the erythrogram while leucogram showed leucocytosis accompanied with heterophilia, lymphopenia and monocytopenia at the 3rd and 7th days post infection. Serum biochemical analysis showed significant elevation in the activity of AST, ALT and AP and in blood urea nitrogen and creatinine concentration along the experimental period. Serum total proteins, albumin, :, ; and < globulin significantly increased at different periods of the experiment. BHV-1 antibodies were detected in the sera of infected rabbits by Dot ELISA and ELISA from the first week until the forth week post infection. Histopathological examination revealed that the most affected organs were the trachea, lungs and liver while adrenals, kidneys, and spleen showed mild pathological alterations.