Veterinary Research Forum

Abstract Numerous studies have explored the molecular epidemiology of H9N2 viruses in Iran; however, continuous monitoring remains vital for timely interventions to mitigate potential damage. This study examined the molecular characteristics and evolutionary features of Iranian H9N2 viruses by sequencing the complete genomes of two viruses, Marand and Baneh, isolated in 1998 and 2022, respectively, alongside other Iranian strains from GenBank. All Iranian viruses were identified as low-pathogenic avian influenza viruses, as evidenced by the presence of the di-basic motif K/RSSR cleavage site. Notably, all Iranian viruses isolated from 2009 onward had an L at position 216 in the hemagglutinin receptor binding site, whereas earlier viruses exhibited a Q/L at the same position, an essential mutation that enhances replication in mammalian cells. The molecular evolutionary rates for the Iranian hemagglutinin (HA) and neuraminidase (NA) genes were estimated at 4.50 × 10^-3 and 3.60 × 10^-3 substitutions per site per year, respectively. Error-prone replication of H9N2 viruses has resulted in the continuous evolution of Iranian strains over two decades, characterized by three phases of population growth. Maximum likelihood phylogenetic analysis revealed that the HA and NA genes of H9N2 viruses from domestic chickens belonged to the G1 sublineage. Additionally, the internal genes of some viruses displayed evidence of reassortment with other subtypes, indicating potential gene exchange with other viruses. These findings underscored the importance of ongoing surveillance of H9N2 viruses in both domestic and wild bird populations, given the human-like receptor-binding preference and the possibility of genetic reassortment with various viral subtypes.

Abstract Bovine respiratory disease (BRD) is a significant disease in the cattle industry worldwide. The interaction between environmental factors, hosts, livestock management, and viral and bacterial pathogens causes this disease. Viruses are crucial in the initiation and progression of BRD. This study was the first to investigate the prevalence of BRD viruses using the reverse transcription polymerase chain reaction method in nasal and eye conjunctival swabs and blood samples of 115 BRD calves in the central desert of Iran. At least one investigated virus was detected in 44 animals (38.26%). The detection rates of bovine viral diarrhea virus, bovine coronavirus, bovine adenovirus, bovine respiratory syncytial virus, bovine herpes virus-1, and bovine para influenza virus-3 were 20.00, 14.78, 5.21, 0.86, 0.00, and 0.00%, respectively. Three animals (2.60%) had a simultaneous infection with two viruses. Detection of bovine viral diarrhea virus, bovine coronavirus, and bovine adenovirus was correlated. The virus infection rates were 31.81 and 44.66% in five sampled cities. The virus detection rate in infected animals was related to the nose (26 animals; 50.09%), nose and eyes (seven animals; 15.90%), eyes (seven animals; 15.90%), nose, eyes, and blood (three animals; 6.81%), and blood (one animal; 2.27%) samples. The virus detection rate in different samples was in separate clusters. Monitoring and controlling the circulation of bovine viral diarrhea virus and bovine coronavirus in the central desert of Iran is vital due to the high detection rate. Our results highlight the necessity of investigating other viruses and bacterial agents related to the BRD in the study area.

Abstract Trichinella britovi is a species circulating in Iran and has already been observed in wild and domestic carnivores. The present study aims to determine the occurrence of Trichinella species in Iran's most important provinces, including the northern, northeastern, and northwest regions. In total, 93 carcasses of golden jackals, collected due to road accidents between 2019 and 2022, have been obtained from Ardabil, Mazandaran, Golestan, North Khorasan, and Khorasan Razavi provinces, Iran. Genomic DNA (four positives out of 93 carcasses) from each Trichinella genotype was amplified in a multiplex polymerase chain reaction (PCR) reaction. To confirm the multiplex PCR results, the sequences of the PCR products with cytochrome oxidase subunit I were sent to a reference laboratory within the European Union. Bioinformatics software was used to verify the sequencing results of primer fragments and to compare and analyze the results obtained with the sequences stored in the NCBI genetic database. Following multiplex PCR amplification, a two-band pattern was observed. Trichinella britovi larvae displayed the expected band pattern of 127 bp and 253 bp. The PCR products amplified using primers specific for the cytochrome oxidase subunit I gene showed 861 base pair bands in all four samples analyzed. The obtained sequences of cytochrome oxidase subunit I from all four individual muscle larvae had 100% identical haplotypes except for one nucleotide transversion in one sequence. As a result of this study, valuable insights into T. britovi prevalence and phylogenetic distribution among Iranian golden jackals have been gained in terms of prevalence and phylogenic distribution.

Abstract Bovine ephemeral fever (BEF) is a debilitating disease of cattle and water buffaloes. Bovine ephemeral fever viruses (BEFVs) form four phylogenetic lineages including the Middle East, East of Asia, Australia and Africa, while the exotic viral strains have also been detected in different geographic areas. We characterized eight BEFVs from different regions of Iran during a period of seven years from 2015 to 2022. Sequencing the entire length of the G gene, the BEFVs were classified in the Middle Eastern lineage with the maximum of 99.73% and minimum of 97.30% nucleotide identity. The all Iranian and Turkish BEFVs detected during the large epizootic in 2020 were clustered phylogenetically together. However, no amino acid variation was observed between the Iranian viruses detected in 2020 and those identified before 2020 in the Middle Eastern lineage suggesting that host, environmental and other genetic factor (s) might have involved in occurrence of the epizootic in 2020. Two BEFVs detected during 2022 outbreak from Kermanshah and Narmashir in the west and east of Iran, respectively, were clustered in two distinct groups as a novel amino acid substitution H51Y in the epitope G3 was also identified in Kermanshah 2022 sequence. These results imply that the Middle Eastern lineage replaced the previously circulated East Asian BEFVs in Iran during 2012 to 2013 and also signify the emergence of new BEFVs due to the intra-lineage evolution. Continuous monitoring of the circulating viruses and identifying the potential vector (s) and its biology help better understand epizootiology of BEFV in the high-risk region.

Abstract Dogs serve as definitive or intermediate hosts for some protozoan and helminth parasites, some of which are of zoonotic, and others cause severe infectious diseases in other animals. The aim of this study was to determine the prevalence of intestinal parasites in shelter dogs in Khorasan Razavi province, Iran. A total of 323 fecal samples were randomly collected from shelter dogs in Mashhad and Neyshabur areas. All samples were then transferred to the Parasitology Laboratory and examined detect intestinal protozoa and helminths using the formalin-ether technique. Out of the 323 fecal samples, 92 (28.48%) tested positive for intestinal parasites. The overall prevalence of intestinal parasites in shelter dogs in Mashhad and Neyshabur areas were 23.83 and 35.38%, respectively. Monospecific infection was found in 19.50% of dogs, while concurrent infection with two or more species was seen in 8.97% of dogs. The frequencies of the nine detected parasites were as follows: Sarcocystis sp. 12.38%, Giardia sp. 4.64%, Cystoisospora sp. 2.78%, Hammondia Neospora-like oocysts 2.16%, Taenia sp. 6.19%, Toxascaris leonina 7.43%, Dicrocoelium dendriticum 1.54%, Toxocara canis 0.92% and Capillaria sp. 0.30%. The results of this study showed the high frequency of intestinal parasites in shelter dogs. To control important zoonotic helminthic diseases such as echinococcosis and toxocariasis, it is recommended to eliminate stray dogs in cooperation with municipalities and veterinary departments in Iran. If stray dogs are transferred to shelters, they should receive regular treatment with effective anti-helminthic drugs.

Abstract Crimean-Congo hemorrhagic fever (CCHF) is indeed to be considered as one of the most significant vector-borne diseases globally. The virus responsible for CCHF can persist in various animals and lead to severe infections in humans. Ticks of the Ixodidae family are the acknowledged vectors of CCHF virus (CCHFV) transmission to humans. In this review, different tick species from Iran and its neighbors and their roles in the transmission of CCHFV were evaluated. However, the transmission patterns of CCHF to humans primarily involve tick bites, direct contact with the blood, organs, or tissues of contaminated animals, and exposure to CCHF patients are important in virus transmission. The results of this review confirm that the tick vectors in Iran and its neighbors are very common. Even though tick bite is one of the most important risk factors for CCHF, it cannot explain all cases, and there are other important risk factors, such as high-risk occupations and having contact with livestock. Ticks that have been proven to transmit the disease, especially Hyalomma marginatum, are scattered in Iran and neighboring countries. Transporting passengers and livestock and sometimes smuggling livestock are the means of transmitting the virus between neighboring countries. Therefore, there is a need for joint preparedness and response programs to prevent and manage CCHF between Iran and its neighbors.

Abstract Hydropericardium syndrome (HPS) has caused significant financial losses to the Iranian poultry industry in the past few years. Thirty-two broiler chickens with gross lesions of HPS were inspected histologically and immunohistochemically. Sampling was performed in Sabzevar, Iran. The dead and sick birds from random farms were subjected to necropsy examinations. Only four broiler chickens had no hydropericardium and the other gross findings were similar for birds. Basophilic and eosinophilic intranuclear inclusions, hemorrhages and necrosis in different organs were the primary characteristic histologic lesions. Lymphoid depletion, goblet cell hyperplasia and necrotizing enteritis were some of the findings reported in previous research. Low macrophage infiltration rate and brain lesions were other discoveries in Hematoxylin and Eosin (H&E) examination. Feulgen reaction and Cluster of Differentiation 68 (CD68) immunohistochemical staining were used for a comprehensive investigation and these techniques revealed improved histopathologic details. Feulgen staining confirmed brain lesions and some other changes in different organs. Eventually, the CD68 method revealed low macrophage presence in most organs. This study suggested that HPS might cause brain damage and the susceptibility of the Arian breed to the adenovirus needs further investigation.

Abstract A 6-year-old male domestic short-haired cat (Felis catus) was presented with weakness, anorexia, fever, icterus, a painful abdomen, ruffled hair and a tick infestation, and it had no prior surgery. Laboratory analysis revealed left-shifted neutrophils, thrombocytopenia, low albumin content and high serum bilirubin concentration as well as activities of hepatic enzymes including alanine aminotransferase and aspartate aminotransferase. Azotemia and increased serum levels of creatinine and urea were also recorded. In Giemsa-stained blood smear, Hepatozoon gamonts were observed within neutrophils. Species-specific polymerase chain reaction assay was used to amplify an approximately 590 bp fragment of 18S rRNA gene and confirmed Hepatozoon felis infection. The cat was treated with imidocarb dipropionate and doxycycline and recovered completely. Six-month follow-up showed no recurrence. This study reveals the presence of H. felis in Iran and it should be considered in differential diagnosis in febrile and icteric cats. To the authors' knowledge, this is the first description of H. felis infection in a cat in Iran.

Abstract Giardia duodenalis is a zoonotic protozoan infecting various vertebrates such as humans and domestic animals. The aim of this study was to determine the frequency and genotypes of G. duodenalis using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in dogs of Urmia, Iran. Overall, 246 stool specimens were collected from 100 pet, 49 stray, and 97 shelter dogs in the Urmia, Iran. Totally, seven samples (2.48%) were microscopically positive in terms of Giardia cyst. The PCR-RFLP analysis revealed that three (1.21%) and two (0.83%) samples have the C and D genotypes, respectively. In addition, two samples (0.83%) were belonged to the AI sub-group. A significant association was determined between the frequency of Giardia infection and life style, age, and stool form of dogs. The findings of the study showed the high frequency of Giardia infection in stray dogs and the dogs under one-year-old. Furthermore, the C and D genotypes of G. duodenalis were predominant in dogs of Urmia, Iran.

Abstract The present study reported the first serotyping (O:H typing) data documented in Shiga toxin-producing Escherichia coli (STEC) strains of animal origin in Iran in isolates recovered between 2008 to 2016. A total number of 75 STEC strains previously isolated from fecal samples of cattle, sheep, goats, pigeons, humans, and deer were assessed by different polymerase chain reaction (PCR) assays detecting the major virulence genes of STEC and phylogroups. Then, the strains were tested for the 16 important O-groups by PCR. Finally, twenty strains were selected for H-genotyping by PCR plus sequencing. The predominant serogroup was O113 which was detected in nine isolates (five cattle, 55.50%; two goats, 22.20%; two red deer, 22.20%) followed by O26 (3/3, 100%) in cattle, O111 (3/3, 100%) in cattle, O5 (3/3, 100%) in sheep, O63 (1/1, 100%) in pigeon, O75 (2/2, 100%) in pigeons, and O128 in goats (2/3, 66.60%) and pigeon (1/3, 33.30%). The most important recognized serotypes were O113:H21 in cattle (2/3) and goat (1/3), O113:H4 in red deer (1/1), O111:H8 in calves (2/2), O26:H11 in calve (1/1), O128:H2 in goats (2/3) and pigeon (1/3), and O5:H19 in sheep (3/3). One cattle strain carrying stx1, stx2, eae, and Ehly genes belonged to O26:H29 serotype. Most strains with determined O-groups were from the bovine source that highlighted the importance of cattle as reservoirs of potentially pathogenic serovars. The present study suggested that the top seven non-O157 serogroups should be assessed along with O157 in all future research and clinical diagnostics of STEC in Iran.

Abstract There are few studies on Coxiella burnetii (Cb) as a causative agent of Q fever in dairy products in Iran. The prevalence of Cb was studied by polymerase chain reaction (PCR) method in Kope (pot) cheese and cattle milk collected from West Azerbaijan province, Iran. A total number of 240 Kope cheese and 560 milk samples were collected during the year 2020. All samples were subjected to PCR based on transposable gene IS1111. The results showed that 12.50% (95.00% confidence interval (CI): 9.00% - 16.10%) of Kope cheese and 13% (95.00% CI: 10.00% - 17.30%) of milk samples were positive for Cb. There was a significant difference in cheese and milk contaminations with Cb among the defined age groups as well as regional and seasonal variations. It was concluded that Kope cheese and cattle milk are important sources of Cb and should be considered as important risk factors in the epidemiology of Q fever disease in public health.

Abstract Leishmaniasis is a protozoal and vector-borne disease. World health organization has considered the disease as a neglected tropical disease. Phlebotomus and Lutzumyia species (order: Diptera, family: Psychodidae) are human leishmaniasis vectors in new and old worlds. Sergentomyia spp. (Diptera, Psychodidae) are proven vectors of lizard leishmaniasis. Although some studies have identified human Leishmania parasites in Sergentomyia, their role in parasite circulation is unknown yet. Hence, the parasitological and molecular methods were used to study the possible Leishmania infection of Sergentomyia spp., in the human and canine visceral leishmaniasis endemic area in North West of Iran. Even though Sergentomyia specimens were caught in a dominant number compared to Phlebotomus spp., no Leishmania promastigote or DNA was detected in live-caught or sticky trap-caught specimens, respectively. Sergentomyia spp. are proven vectors of sauroleishmaniasis, and despite several global reports of Leishmania infection in Sergentomyia spp., such findings should be carefully interpreted to avoid false vector incriminations.

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 Kobuviruses are viral pathogens with broad host range presented in human gastroenteritis cases; but, the pathogenesis of these viruses in companion animals is not well described. In the present study, the presence of canine (CaKVs) and feline kobuviruses (FeKVs) was detected in the 100 fecal samples of diarrhoeic and healthy companion dogs and cats by polymerase chain reaction in Tehran, Iran. The prevalence of infection was estimated as 8.00% and 4.00% in dogs and cats, respectively. All positive samples were belonged to non-diarrhoeic animals except for a feline sample being co-infected with panleukopenia. Sequence analysis showed multiple point mutations in canine and feline Iranian strains and new feline strain was detected in the present study. This is the first detection of CaKVs and FeKVs in Iran; but, the exact role of these enteric viral pathogens and their zoonotic risks are better to be clarified in all endemic regions.

Abstract A 2-year-old female Shih Tzu dog was submitted with the history of anorexia and depression for one week and no prior surgery. Fever and pale mucosa were noticed in physical examination. Microscopic examination of the Giemsa-stained blood smear disclosed large form of Babesia and single to four pear-shaped merozoites within erythrocytes (RBCs). Regenerative anemia characterized by a marked reticulocytosis, significant intra-vascular hemolysis, nucleated RBCs, left-shifted neutrophils, thrombocytopenia, azotemia, high serum creatinine and urea concentrations were recorded following hemato-biochemical analysis. Abundant bilirubin crystals and abnormal reddish color after centrifugation were observed in urinalysis. Molecular analysis was performed using specific primers for detection of Babesia canis. Diminazene aceturate, ciprofloxacin, ivermectin and phosphorus-vitamin B12 were prescribed and the clinical signs improved after four days. Two months follow-up showed no recurrence. Such studies would significantly contribute to the development of appropriate preventive strategies and successful treatment. This communication reports a clinical case of canine babesiosis caused by B. canis in a female Shih Tzu dog.

Abstract 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.

Abstract Parasitic nematodes of ovine abomasum are of economic and hygienic importance throughout the world and Iran. This study was aimed to evaluate molecular identity and species diversity of parasitic nematodes in the abomasum of slaughtered sheep in Ilam, Iran. In this study, a total number of 240 of abomasa were randomly collected from the slaughtered sheep at industrial slaughterhouses in Ilam in all seasons between 2017 and 2018. The abomasum content and abomasal mucosa were removed and washed. The collected nematodes were morphologically identified. The genomic DNA was extracted and a 300 bp-fragment-length from internal transcribed spacer 2 ribosomal ribonucleic acid (ITS2-rRNA) gene was amplified. Overall prevalence was 66.70% (160/240). Five species of four genera of nematodes including Marshallagia marshalli (43.70%), Ostertagia circumcincta (15.50%), Parabronema skrjabini (5.00%), M. occidentalis (2.50%), and Haemonchus contortus (0.04%) were identified. Ostertagia circumcincta and H. contortus were found to be different in two nucleotides. There was no nucleotide difference between M. marshalli and M. occidentalis. This study revealed a significant prevalence of parasitic nematodes in sheep abomasum and species diversity of Trichostrongylid nematodes in the region.

Abstract Spondylolisthesis known as kinky back syndrome is a deformity of the free sixth thoracic vertebra including enlargement and rotation resulting in posterior paralysis in broiler chicken. Genotype, feeding regimes and environment are involved in occurrence of this congenital anomaly. Spondylolisthesis may result in or happen simultaneously with two other major axial skeleton pathologies including spondylosis and spondylitis as a result of occurrence of degenerative necrobiotic lesions and inflammation of the vertebrae, respectively. This report deals with a male broiler chicken with posterior paralysis in the second week of rearing. After euthanasia, the specimen was evaluated at macroscopic and microscopic levels. Macroscopic findings showed the unilateral enlargement of body and transverse process of sixth thoracic vertebra and downward rotation of the body. The histopathological studies revealed the focal chondronecrosis and degenerative changes in articular facets and cranial process of the vertebra. In conclusion, a diagnosis of spondylolisthesis followed by spondylosis was made based on macroscopic and histopathological findings that to the authors’ best knowledge, is the first report in a broiler farm in Iran.

Abstract Over the last decade, diagnostic tools to detect and differentiate Fasciola species have improved, but our understanding of the distribution of haplotypes and population structure of this parasite is less clear. This study was designed to survey this gap in the F. gigantica epidemiology in Kermanshah province, western Iran from 2015 to 2017.Sixty-eight Fasciola isolates were collected from slaughterhouses from this province. We evaluated the PCR-RFLP assay of the ITS1 genes for the identification of Fasciola species using the RsaI enzyme. After Fasciola species identification, the partial sequence of mitochondrial NADH dehydrogenase subunit 1 (ND1) gene of F. gigantica was used for subsequent construction of the phylogenetic tree and network analysis.Based on the PCR-PRFLP profile, one (6.25%) of sheep isolates and 19 (39.60%) of cattle isolates were detected as F. gigantica, whereas 93.75% of sheep isolates, 60.40% of cattle isolates and all of the goat isolates were F. hepatica. In the 20 analyzed flukes, five ND1 haplotypes were detected. Statistically significant genetic differentiation was demonstrated between the Iran population and all the other populations. Evidence is presented for the existence of two well-separated populations: African and West Asian gigantica flukes and East Asian gigantica flukes.Genetic relationships among haplotypes were associated with geographical divisions. Also, our results have heightened our knowledge about the genetic diversity of F. gigantic, providing the first evidence for the existence of two well-separated populations of this parasite.

Abstract Various species of Trypanosoma parasites are known to infect several wild and domestic animals worldwide. A 7-year-old Holstein cow from Baneh, Kurdistan province, was examined by a private veterinarian due to anorexia and depression. Physical examination revealed fever, enlarged subscapular lymph node, and pale mucosa. Blood samples were taken for hemato-logical, parasitological, and PCR examination. The large Trypanosoma spp. was microscopically observed in a stained blood smear. Decreased red blood cells (RBCs) count, packed cell volume and hemoglobin concentration were observed through complete blood cell count. Nucleated RBCs were also found in this case. Species-specific PCR assay confirmed T. theileri infection. Treatment was performed subcutaneously with diminazene aceturate. The clinical signs were improved after two days. Two-month follow-up showed no recurrence. In conclusion, T. theileri is characterized by anemia and pyrexia in a cow. To our knowledge, the present case report describes the first molecular evidence of T. theileri in Kurdistan, West of Iran.

Abstract Razi Vaccine and Serum Research Institute (RVSRI) turned 95 years old in 2015. Majority of the animal infectious diseases such as rinderpest and anthrax that used to frequently strike the historic Persia are now gone for good or under control owing to the pioneering researches conducted at the institute in the early-mid 20^th century in the field of vaccine manufacturing. The earliest such scientific contributions, were truly made by the French eminent veterinarian Dr. Louis Pierre Joseph Delpy who joined the institute in 1931. In his 18 year-long directorship tenure he taught his colleagues fundamentals of vaccinology, basics of modern epidemiology, essentials of infectious disease control disciplines, the art of scientific writing and much more things that changed the institute for ever. This paper reviews the events and turning points in the first 25 years of service of the institute in a chronological way and remarks Delpy’s principle involvements in all of these on the occasion of the 120 anniversary of his birth. At the entrance of the institute headquarter building where his bronze bust is placed, visitors can see a memorial etched plate that reads “... The architect of Razi and founder of Archives De L’Institute Razi (Archives of Razi Institute) was an enthusiastic scientist with a creative mind. … For the Razi community, Dr Delpy is gone but not forgotten.”

Abstract Influenza is a highly contagious zoonotic disease in the world. Avian H9N2 influenza virus is a significant pandemic pathogen widely distributed throughout the world. Pet ownership has been documented as a risk factor for infection transmission to human. Considering major public health concern, the prevalence of antibodies against avian H9N2 influenza virus was evaluated in 170 serum samples of dogs by hemagglutination inhibition assay. This study is the first survey to assess the epidemiology of avian H9N2 influenza virus infection in dogs in Kerman, southeast of Iran. Out of 170 samples, 65 (38.23%) were positive for H9N2. Antibodies were higher in farm dogs that were kept with other animals and also in dogs were fed a raw diet. These findings emphasize the importance of close attention to these populations for control and prevention programs. It is important to reduce infection burden, especially in regions with widespread distribution of H9N2.

Abstract Genus Brachyspira,as Gram negative anaerobic bacteria, colonize in dogs intestine. The aim of the current study was to determine the prevalence of Brachyspira spp. for the first time in Iran and rapid identification of Brachyspira spp. in dogs by a new designment of a species-specific primer set for B. canis. One hundred fifty-one fecal samples were obtained from dogs by rectal swab. Twenty dogs suffered from diarrhea and 131 of them were healthy. In 9.27% (14/151) of samples, spirochaetes were detected on primary cultures by weak hemolysis and positive Gram staining and then Brachyspira genus was confirmed by NADH oxidase (nox) gene via polymerase chain reaction. Among 14 isolates, twelve isolates were B. canis, one isolate was B. intermedia and another one was non-typeable. From 12 B. canis, only eight isolates were detected by designed specific primers. Ten Brachyspira spp. were isolated from dogs ≤ 1 year old (10/67, 14.92%) and 4 isolates were from > 1 year old dogs (4/84, 4.76%). The isolation rates from healthy and diarrheic dogs were (12/131, 9.16%) and (2/20, 10.00%), respectively. A statistically significant association was observed between the presence of Brachyspira spp. and the age under one year. Based on our findings, the nox gene in B. canis might have more sequence variability compared to other Brachyspira spp.

Abstract A polycephalic larva of Taeniidae family isolated from abdominal cavity of a great gerbil (Rhombomys opimus) from Golestan province, northern Iran, was subjected to molecular analysis. Genomic DNA from the larva was obtained using a DNA extraction tissue kit. Polymerase chain reaction was performed for amplification of the partial 12S rRNA, cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase 1 (nad1) mitochondrial genes. BLAST analysis of DNA sequencing indicated 99.00% homology in 12S rRNA and cox1 genes and 98.00% homology in nad1 gene with Hydatigera krepkogorski (accession No. AB731762). The sequences of current isolate were deposited in GenBank by accession Nos. MF281971, MF281972 and MF281973 for 12 SrRNA, cox1 and nad1 genes, respectively. This study was the first report of molecular characterization of H. krepkogorski from Iran. Isolation and characterization of the adult stage from definitive host will help to better clarify incomplete life cycle and morphology data of this species in the world.

Abstract Fungal infections cause significant economic losses in the poultry industry either due to their direct infectious nature or due to mycotoxins production. Hatchery contamination with fungi can threaten chicken health. In this regard, geographical and seasonal distributions of airborne fungal contamination of 25 hatcheries in Mazandaran province, northern Iran, were investigated using an open plate method. The results of this study showed that hatcheries have various fungal contaminations, among which the most common were respectively Cladosporium (31.07%), Penicillium (24.00%), Aspergillus (20.63%), sterile hyphae (14.70%) and Alternaria (6.20%) from different regions. The results revealed that the highest level of fungal isolation was in spring and autumn. This study also showed that the concentration of fungal air spora in forest and seaside locations was significantly greater than mountainous ones. In spite of the regular disinfection in commercial hatcheries, fungal contamination was found in different parts.