Investigation of pyrethroid resistance mutations in Linognathus stenopsis lice collected from goats in western and northwestern Iran.

Introduction: Linognathus stenopsis lice are an extensive parasitic concern in goat populations worldwide, posing significant economic and health risks. This study examined the identification of alleles of resistance to pyrethroid and mutations in L. stenopsis samples obtained from goats in five provinces in western and northwestern Iran. Methods: Morphological and molecular techniques were employed to identify the louse species. Molecular identification methods and gene sequencing were used to identify resistance-associated mutations in the voltage-gated sodium channel (VGSC) gene. Results and discussion: The results revealed that six amino acid substitutions, including threonine-to-isoleucine (T917I), leucine-to-phenylalanine (L920F), isoleucine-to-phenylalanine (I927F), phenylalanine-to-alanine (F928A), valine-to-arginine (V929R), and arginine-to-leucine (R930L) mutations, were present in the VGSC gene of L. stenopsis lice from various regions of Iran. These findings suggest the potential for pyrethroid resistance development in this louse species, highlighting the importance of integrated pest management (IPM) strategies. Such strategies, which combine selective insecticides, regular grooming, and environmental sanitation, are crucial for effectively managing L. stenopsis infestations and preserving the efficacy of pyrethroids for pest control. Moreover, the emergence of novel kdr mutations underscores the need for ongoing research into the molecular mechanisms underlying these mutations. This research is vital for developing strategies to combat pyrethroid resistance and maintaining the efficacy of insecticides in controlling lice.

Adding yeast extract to culture medium enhances replication of the avian coronavirus infectious bronchitis virus in chicken embryo kidney cells.

Infectious bronchitis virus (IBV), an avian coronavirus, can be isolated and cultured in tracheal organ cultures (TOCs), embryonated eggs and cell cultures, the first two of which are commonly used for viral isolation. Previous studies have suggested that foetal bovine serum (FBS) can inhibit coronavirus replication in cell cultures. In this study, the replication of IBV in chicken embryo kidney (CEK) cell cultures and the Leghorn hepatocellular carcinoma (LMH) cell line was assessed using two different cell culture media containing FBS or yeast extract (YE) and two different IBV strains. The highest concentrations of viral genomes were observed when the cell culture medium (CEK) contained YE. Similar results were observed in LMH cells. Examination of the infectivity by titration demonstrated that the cell lysate from CEK cell cultures in a medium including YE contained a higher median embryo infectious dose than that from CEK cell cultures in a medium containing FBS. These results indicate that improved replication of IBV in cell cultures can be achieved by replacing FBS with YE in the cell culture medium.

Detection of pyrethroids resistance alleles in goat biting louse Bovicola caprae (Phthiraptera: Trichodectidae) in west and northwest of Iran.

Pyrethroid insecticides target voltage-gated sodium channels (VGSCs) that are essential for electrical signaling in the nervous system of insects. Three-point mutations at the corresponding amino acid sequence positions M815I, T917I, and L920F located in domain II conferring the knockdown resistance (kdr) are the most important mutations in pyrethroid-resistant lice worldwide. In addition, six new mutations have been reported in the extracellular loops IIS1-2 (H813P) and IIS5 (I927F, L928A, R929V, L930M, L932M) in the α- subunit of the sodium channel in lice. The aim of this study was to detect alleles resistant to pyrethroids in the domain II (S5-S6) of the VGSC gene in goat biting louse. Goat biting lice were collected from five provinces in the west and northwest of Iran. Genomic DNA was extracted from goat biting lice and Bovicola (Damalinia) caprae species was confirmed by amplifying the cytochrome oxidase subunit I (COXI) gene. A fragment in the domain II (S5-S6) of the VGSC gene was amplified using the specific primers and the resultant polymerase chain reaction products were sequenced. Substitutions T917I, L920F, I927F, L928A, R929V and L930M were identified in the examined sequences. The results showed that all the examined lice had at least one mutation in their VGSC gene associated with pyrethroid resistance or new mutations. The presence of these mutated alleles in the VGSC gene may be due to the long-term and multiple use of pyrethroids against arthropods. Thus, the molecular detection of resistance to pyrethroid insecticides in goat chewing lice can help plot a kdr frequency map to enact effective policies to control caprine pediculosis.

Molecular study on nematode infection in sheep abomasa: a regional investigation in Iran and Iraq.

Gastrointestinal nematodes are leading causes of loss in livestock and are the primary restriction to its profitable production, worldwide. This study was carried out to determine the prevalence and diversity of sheep abomasum nematode species in Urmia (Iran) and Soran (Iraq) slaughterhouses from October 2019 to January 2021. A total of 280 abomasa (each city 140 samples) were randomly collected from the slaughtered sheep. The abomasal content and mucosa were removed and washed. The collected nematodes were morphologically identified. Genomic DNA was extracted from identified nematodes and a fragment from the internal transcribed spacer 2 ribosomal ribonucleic acid (ITS2-rDNA) gene was amplified. In Urmia city, two species including Teladorsagia circumcincta (40.7%), and two morphotypes of Marshallagia species; Marshallagia marshalli (35.0%) and M. trifida (4.3%) were identified. In Urmia city, 52.9% of the examined sheep were infected with at least one species of nematodes. The overall prevalence of abomasa infection with nematodes in Soran city was 91.4%. In the examined sheep abomasa in Soran city, four species of nematodes were identified, including Marshallagia species with two morphotypes, M. marshalli (85.0%) and M. trifida (20.7%), Teladorsagia circumcincta (32.1%), Parabronema skrjabini (1.4%), and Haemonchus contortus (0.7%). Except for H. contortus, all the other identified nematode species were confirmed using molecular techniques. It was concluded that abomasal nematode infections are widespread in sheep particularly in Soran city. Marshallagia marshalli and T. circumcincta were most prevalent nematodes in both regions. In addition, further molecular studies are recommended to understand the intra-specific variations in the genus Marshallagia and more accurate identification of morphotypes in these regions.

Phylogenetic and molecular analysis based on genes 16S-rRNA, OMPA and POMP to identify Chlamydia abortus infection occurrence at the milk samples of goats and sheep in west Azerbaijan of Iran.

BACKGROUND AND OBJECTIVES: Enzootic abortion in sheep and goats, also called ovine enzootic abortion (OEA) or enzootic abortion of ewes (EAE), is caused by Chlamydia abortus. The disease has a major economic impact as it represents the most important cause of lamb loss in sheep in parts of Europe, North America and Africa. This serious and potentially life-threatening zoonosis can also affect pregnant women after contact with lambing ewes, leading to severe febrile illness in pregnancy and loss of the foetus. MATERIALS AND METHODS: The present study was conducted to the Phylogenetic and Molecular Analysis based on Genes 16S-rRNA, OmpA and POMP of C. abortus in milk samples collected from sheep and goats in West Azerbaijan province, Iran. During 2018, a total number of 360 milk samples were collected from sheep (n = 180) and goats (n = 180) of different regions of the province. All milk samples were subjected to DNA extraction and examined by PCR. RESULTS: Among 360 milk samples collected from sheep and goats, 31 (8.611%; 95% CI=6.13-11.96) were positive for Chlamydia spp. The helicase, 16S-rRNA and ompA genes were examined and resulted in 8, 31, 31 of positive samples respectively. The accession numbers have been deposited in GenBank (NCBI) (MT367602 and MT367603). CONCLUSION: Phylogenetic analysis based on the gene of helicase showed that most of the isolates shared similarity > 99.97%.

Changes in rainbow trout (Oncorhynchus mykiss) growth and mucosal immune parameters after dietary administration of grape (Vitis vinifera) seed extract.

The effect of dietary grape (Vitis vinifera) seed extract (GSE) on growth performance and mucosal immune parameters in rainbow trout (Oncorhynchus mykiss) fry was studied. Fish (1.3 g mean weight) were randomly distributed in nine tanks (15 fish per tank) and fed diets containing GSE at 0 (control), 100, and 200 mg kg-1for 60 days. The results showed that growth parameters were enhanced in both treatment groups compared to the control group. Histological examination of fish skin showed higher epidermis thickness, goblet cell density, and volume density in the GSE groups compared to the values of the control group. Furthermore, the villus height, goblet cell density, and intraepithelial lymphocytes were increased in the fish intestine in those fish fed GSE, with respect to control fish. Feeding fish with low dose of GSE (100 mg kg-1) up-regulated the expression of some immune-relevant genes, including complement component 3 (C3), lysozyme (Lys), omDB-3, interferon gamma (IFN-γ), and tumor necrosis factor-α (TNF-α) in different mucosal tissues. However, feeding fish the high dose of GSE (200 mg kg-1) mostly enhanced expression of these genes in the skin. Besides, skin mucus of fish fed GSE showed bactericidal activity against Yersinia ruckeri. It was concluded that GSE, especially at 100 mg kg-1, modulates the growth performance and mucosal immunity of rainbow trout.

Molecular detection, phylogenetic analysis, and antibacterial performance of lactic acid bacteria isolated from traditional cheeses, North-West Iran.

Lactic acid bacteria (LAB) are candidate probiotic bacteria that can provide health benefits when delivered via functional foods. The purpose of this study was to isolate and characterize LAB from traditional cheeses consumed in north-west regions of Iran. A number of sixty traditional cheeses samples were collected and initially screened as LAB using biochemical and molecular methods. A fragment of 1,540 bp in size of 16s rRNA gene was amplified from 70 bacterial isolates. Restriction fragment length polymorphism (RFLP) was employed to differentiate LAB isolates. LAB isolates generated three different RFLP patterns using HinfI restriction enzyme. Phylogenetic analysis revealed that LAB isolates belonged to three genera including Enterococcus, Lactobacillus, and Lactococcus. Most of the isolated LAB strains belonged to Enterococcus spp. The antimicrobial performance of eight LAB isolates with different RFLP patterns ranged from 6.72 to 14.00 mm. It was concluded that molecular characterization of LAB strains in traditional cheeses will enhance our understanding of traditional food microbiota and will help to find bacterial strains with probiotic potential with great benefit both in health and industry.

Molecular analysis of pyrethroid resistance in Cimex hemipterus (Hemiptera: Cimicidae) collected from different parts of Iran.

The present study was aimed to assess the bedbugs susceptibility to pyrethroid insecticides using molecular analysis. With the aid of pest control companies, adult bedbugs were collected from various places such as hotels, residential houses, and industrial buildings in seven cities highly crowded with domestic and foreign tourists in Iran from May 2016 to August 2017. Bedbugs were colonized in the laboratory to evaluate their resistance to pyrethroid using insecticide resistance bioassay. Genomic DNA was extracted from susceptible and resistant bedbugs. At first, specie specific primers targeting cytochrome oxidase subunit I (COI) gene was performed to confirm Cimex hemipterus species. Then, kdr-like gene was examined for point mutation using PCR and nucleotide sequencing. Bioassay showed that 11 out of 35 examined bedbugs were resistant to pyrethroids (31.43%; 95.00% confidence interval: 29.48-33.08%). The DNA sequencing showed that all examined bedbugs collected from Tehran province had homozygous V419L kdr-like gene mutations. The level of pyrethroid resistance found in the collected bugs from Tehran province indicated that this phenomenon has already been prevailed in the site and prompts the need to reevaluate the large use of pyrethroids to control the bedbugs.

Administration of grape (Vitis vinifera) seed extract to rainbow trout (Oncorhynchus mykiss) modulates growth performance, some biochemical parameters, and antioxidant-relevant gene expression.

Grape seed, as a main source of polyphenols, has many nutritional and medicinal properties in humans. In the current study, the effects of dietary ethanolic grape seed extract (GSE) on the growth performance, antioxidant activity, and some biochemical parameters in rainbow trout were investigated. Ninety fish (initial weight 78.47 g) were randomly distributed among nine cement tanks (1.8 m × 0.22 m × 0.35 m) with 10 fish per tank. Three experimental diets containing either 0, 10, or 50 g kg-1 GSE were prepared and each diet was randomly assigned to three tanks of fish for 60 days. Results showed that feeding GSE enhanced some growth parameters including the specific growth rate and condition factor in comparison with the control group. Among different serum metabolites, the glucose levels in treatment groups significantly decreased compared to the control group. The total product of lipid peroxidation indicated as malondialdehyde significantly decreased in both the GSE-added treatment groups. The gene expression related to the antioxidant enzymes, catalase, glutathione peroxidase 1, and glutathione S-transferase A, were upregulated in the intestine of fish that received a low dose of GSE. The results of the current study suggest that GSE, especially at 10 g kg-1, diet had the potential to improve (1) specific growth rate and condition factor, (2) biochemical parameters including glucose and lipid peroxidation product, and (3) upregulated the expression of antioxidant genes including catalase, glutathione peroxidase 1, and glutathione S-transferase A in rainbow trout.

The Phylogenetic Analysis of Cimex hemipterus (Hemiptera: Cimicidae) Isolated from Different Regions of Iran Using Cytochrome Oxidase Subunit I Gene.

BACKGROUND: Bedbugs are blood feeding ectoparasites of humans and several domesticated animals. There are scarcity of information about the bed bugs population throughout Iran and only very limited and local studies are available. The aim of this study is to assess the phylogenetic relationships and nucleotide diversity using partial sequences of cytochrome oxidase I gene (COI) among the populations of tropical bed bugs inhabiting Iran. METHODS: The bedbugs were collected from cities located in different geographical regions of Iran. After DNA extraction PCR was performed for COI gene using specific primers. Then DNA sequencing was performed on PCR products for the all 15 examined samples. RESULTS: DNA sequencing analysis showed that the all C. hemipterus samples were similar, despite the minor nucleotide variations (within the range of 576 to 697bp) on average between 5 and 10 Single nucleotide polymorphisms (SNPs). Subsequently, the results were compared with the database in gene bank which revealed close similarity and sequence homology with other C. hemipterus from other parts of the world. CONCLUSION: In conclusion, this study has demonstrated the ability of the COI gene to differentiate between the C. hemipterus populations from a few different locations in Iran. The current research is the first report of phylogenetic and genetic species diversity analysis conducted on C. hemipterus in Iran. These results provided basic information for further studies of molecular epidemiology, public health and pest control operators in Iran.

Efficacy of lyophilized cell-free supernatant of Lactobacillus salivarius (Ls-BU2) on Escherichia coli and shelf life of ground beef.

In the present study, the effect of different concentrations of cell-free supernatant (CFS; 10.00 and 35.00 mg g-1) of Lactobacillus salivarius (Ls-BU2) on chemical, microbial and sensorial specifications of ground beef stored under the refrigerated condition was investigated. The antibacterial activity of CFS on Escherichia coli was also assessed. According to agar-disk diffusion method, CFS of Ls-BU2 revealed a promising antibacterial activity against E. coli in culture media compared to CFS of a well-known probiotic (L. acidophilus LA-5). In meat, CFS of Ls-BU2 showed a minimal effective concentration (MEC) of 35.00 mg g-1 on E. coli, while CFS of L. acidophilus represented a MEC of 45.00 mg g-1. The CFS of Ls-BU2 at 35.00 mg g-1 concentration retained psychrophilic counts of meat at a lower value than maximum accepted level (7 log10 CFU g-1). In a similar trend, CFS of Ls-BU2 at 35.00 mg g-1 concentration was also displayed high sensorial scores compared to other CFS-treated samples. In conclusion, we demonstrated that CFS of Ls-BU2 and to some extent CFS of L. acidophilus could act as a safe food additive for the control of bacterial pathogens and to extend the shelf life of ground beef.

Prevalence of Coxiella burnetii in milk collected from buffalo (water buffalo) and cattle dairy farms in Northwest of Iran.

The present study was conducted to determine the prevalence of C. burnetii in raw milk samples collected from water buffalos and cattle in Northwest of Iran (West Azerbaijan Province). A total number of 840 milk samples were randomly collected from buffalos and cattle belonged to three different geographical regions in west Azerbaijan (the map is necessary). The milk samples were collected seasonally during 2018 and the age of animals were recorded. All the milk samples were subjected to DNA extraction. Nested-PCR was used to detect C. burnetii based on the transposable gene IS1111. The results showed that 16.9% (95% CI: 14.5%-19.6%) of the examined milk samples (19.3% buffalo and 14.6% cattle samples) were positive for C. burnetii. There was a significant difference in C. burnetii shedding in milk between different age groups in cattle but not in buffalos (p value <0.05). The shedding of C. burnetii in milk was highly prevalent in summer (31.1%) (p < 0.05, 95% CI: 26.1%-38.4%). There were significant regional and seasonal variations in the prevalence of C. burnetii in the examined milk samples. It was concluded that buffalo population in west Azerbaijan should be considered as an important factor in the epidemiology of Q fever and consequently in public health.

Phylogenetic analysis of the lumpy skin disease viruses in northwest of Iran.

Lumpy skin disease (LSD) is a devastating viral disease of cattle which has recently spread from Africa into the countries of the Middle East. The aim of the present study was to investigate the relationships among lumpy skin disease viruses (LSDV) isolated from different regions of Iran and the origin and spread of these viruses. In this study, a total of 234 blood samples from clinically affected animals from four provinces in the northwest of Iran were screened for LSDV using polymerase chain reaction (PCR). From 80 positive samples for LSDV detected by PCR, the partial P32 gene (759 bp) of 12 isolates were sequenced and phylogenetically analyzed. LSD viruses were grouped in three subclusters with an overall 97.1-100% nucleotide identity. LSDVs isolated from Gilan showed lowest nucleotide identity with the other LSDVs. Four isolates of LSDV including KO-1, EA-1, EA-3, and WA-3 showed 100% similarity with each other and also with the Neethling strain. Phylogenetic analysis indicated that the identified LSDVs were closely related to each other and had high-sequence homology with other LSDV isolates from Africa. It was concluded that LSD outbreak probably occurred in the northwest of Iran by LSDVs entering the country from Iraq and P32 nucleotide sequence information obtained in the present study is a valuable resource in understanding the genetic nature and molecular epidemiology of local LSDV isolates which can be used for future vaccine development based on the circulating strains in the region.

Phylogenetic analysis of Escherichia coli isolated from broilers with colibacillosis based on gyrA gene sequences.

Escherichia coli isolates from chickens with colibacillosis were assigned to phylogenetic groups based on multiplex polymerase chain reaction (PCR) and antibacterial resistance of E. coli belonging to these groups was examined. Furthermore, the gyrA gene of isolates was sequenced and a phylogenetic tree was generated. A total of 84 E. coli isolates were grouped using multiplex PCR of TSPE4.C2, chuA, yjaA, and gadA molecular markers. Four phylogenetic groups were identified with strains divided as follows: 16 in group A (19.05%), 17 in group B1 (20.24%), 23 in group B2 (27.38%), and 28 in group D (33.33%). Escherichia coli isolates belonging to phylogenetic groups B2 and D were resistant to Soltrim and Flumequine unlike the majority of E. coli isolates that belonged to groups A and B1, and which were susceptible to these antibiotics. The phylogenetic results based on gyrA gene sequences from multiplex PCR revealed that E. coli phylogenetic grouping was in accordance with the clusters obtained in the phylogenetic tree. In conclusion, the comparative sequence analysis of gyrA sequences provides a firm framework for an accurate classification of E. coli and related taxa and may constitute a pertinent phylogenetic marker for E. coli.

Les isolats d'Escherichia coli provenant de poulets avec colibacillose ont été assignés à des groupes phylogénétiques sur la base d'une réaction d'amplification en chaine par la polymérase multiplex (ACP) et la résistance antimicrobienne des E. coli appartenant à ces groupes a été examinée. De plus, le gène gyrA des isolats a été séquencé et un arbre phylogénétique a été généré. Un total de 84 isolats a été groupé à l'aide de l'ACP multiplex utilisant les marqueurs moléculaires TSPE4.C2, chuA, yjaA et gadA. Quatre groupes phylogénétiques ont été identifiés et les souches réparties comme suit: 16 dans le groupe A (19,05 %), 17 dans le groupe B1 (20,24 %), 23 dans le groupe B2 (27,38 %), et 28 dans le groupe D (33,33 %). Les isolats d'E. coli appartenant aux groups phylogénétiques B2 et D étaient résistants au Soltrim et à la fluméquine contrairement à la majorité des isolats d'E. coli appartenant aux groupes A et B1, qui étaient sensibles à ces antibiotiques. Les résultats phylogénétiques basés sur les séquences du gène gyrA provenant de l'ACP multiplex ont révélé que le regroupement phylogénétique des E. coli était en accord avec les groupes obtenus dans l'arbre phylogénétique. En conclusion, l'analyse comparative des séquences gyrA fourni un patron solide pour une classification précise des E. coli et taxons associés et pourrait constituer un marqueur phylogénétique pertinent pour les isolats d'E. coli.(Traduit par Docteur Serge Messier).

Infectious hematopoietic necrosis virus (IHNV) outbreak in farmed rainbow trout in Iran: Viral isolation, pathological findings, molecular confirmation, and genetic analysis.

Infectious hematopoietic necrosis virus (IHNV) is the etiological agent of a contagious disease (IHN) mainly in salmonid fish. In the present study, we isolated and identified IHNV in trout fry from Iranian trout farms with unexplained high mortality in 2016. The affected fry showed cumulative mortality of 90% with the gross pathological signs including exophthalmia and hemorrhage of the eye, skin darkening, abdominal distension, ulceration of the snout, and the visceral pallor and yellowish fluid in the intestine. Histopathological examination revealed marked necrosis in the anterior kidney, liver and spleen with the intracytoplasmic inclusion bodies in the liver sections. Also, intranuclear inclusion body and marginated chromatin were observable in the hematopoietic cells of the kidney. The homogenates tissues of infected fry induced IHNV-positive cytopathic effects (CPE) in EPC cells and confirmed by RT-PCR reactions and sequencing. Phylogenetic analysis revealed the Iranian IHNV isolates belonged to the European (E) genogroup with 100% identity to some Italian isolates. This is the first report of IHNV infection in farmed trout fry in Iran describing the viral isolation, clinical symptoms, histopathological findings, molecular confirmation, and genetic analysis suggestion of the specific country of origin.

Prevalence and molecular characterization of staphylococci isolated from sheep with subclinical mastitis in West-Azerbaijan province, Iran.

This study was conducted to investigate the prevalence of subclinical mastitis caused by Staphylococcus spp. in ewes in West-Azerbaijan province of Iran. Molecular characterization of isolated Staphylococcus spp. from diseased ewes were performed using polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) and DNA sequencing of glyceraldehyde-3-phosphate dehydrogenase (gap) gene. Also, antibiotic resistance of staphylococcal isolates against different antibiotics was investigated. A total number of 900 milk samples from 450 native ewes in their mid-lactation period were examined by the California mastitis test (CMT). The CMT positive samples were cultured and bacteria were isolated from 86 (9.50%) glands and 74 (16.40%) ewes. The prevalence of subclinical mastitis in the examined ewes was 16.40%. Microbiological analysis of milk samples revealed that 27 out of 74 sheep with subclinical mastitis were infected with Staphylococcus spp. Amplification of gap gene of 27 Staphylococcus isolates generated a single amplicon of 933 bp in size confirming that isolates were belonged to Staphylococcus genus. Digestion of PCR products by AluI endonuclease generated different RFLP patterns for each species. Nucleotide sequencing of gap gene followed by phylogenetic analysis showed that the most dominant Staphylococcus species were S. epidermidis, S. xylosus and S. chromogenes. Staphylococcal isolates showed the highest resistance to penicillin and ampicillin. In conclusion, Staphylococcus species, except for the southern parts of the province, play an important role in the development of subclinical mastitis in sheep in West-Azerbaijan province of Iran. Also, chloramphenicol, ciprofloxacin and neomycin are the most effective antibiotics for treatment of this disease.

Phylogenetic characterization of virulent Newcastle disease viruses isolated during outbreaks in northwestern Iran in 2010.

The northwest of Iran shares long borders with three neighboring countries; therefore, it is considered one of the main entry portals of Newcastle disease virus (NDV) into the country. Ten virulent NDVs were recovered from 19 poultry farms of various prefectures in northwestern Iran during Newcastle disease outbreaks in 2010. The isolates were genotypically analyzed using an F-gene-specific reverse transcription polymerase chain reaction (RT-PCR) assay. The amplified F gene (nucleotides 189-1666) sequences of the NDV isolates were compared phylogenetically with those of previously published strains in GenBank. All of the NDV isolates belonged to genotype VIIb and were closely related to some isolates from Iran, Russia, and Sweden. Therefore, it can be postulated that these isolates evolved from previously reported strains. The velogenic viruses carried the motif (112)R-R-Q-K-R/F(117) at the F0 cleavage site and a unique substitution of (190)L→F which had never been reported in any NDV genotype VIIb isolate. They shared high sequence similarity with each other but were distinct from current NDV vaccines and NDV strains reported from other countries. This information is fundamental for improving the efficacy of controlling strategies and vaccine development for NDV.

Protective effect of ethyl pyruvate on mice sperm parameters in phenylhydrazine induced hemolytic anemia.

The aim of the present study was to assess the protective effect of ethyl pyruvate (EP) on sperm quality parameters, testosterone level and malondialdehyde (MDA) in phenylhydrazine (PHZ) treated mice. For this purpose, 32 NMRI mice with the age range of 8 to 10 weeks, weight average 26.0 ± 2.0 g, were randomly divided into four equal groups. The control group (1) received normal saline (0. 1 mL per day) by intraperitoneal injection (IP). Group 2 (PHZ group) was treated with initial dose of PHZ (8 mg 100 g(-1), IP) followed by 6 mg 100 g(-1) , IP every 48 hr. Group 3, (Group PHZ+EP) received PHZ (according to the previous prescription) with EP (40 mg kg(-1), daily, IP). Ethyl pyruvate group (4) received only EP (40 mg kg(-1), daily, IP). Treatment period was 35 days. After euthanasia, sperms from caudal region of epididymis were collected and the total mean sperm count, sperm viability, motility and morphology were determined. Testis tissue MDA and serum testosterone levels of all experimental groups were also evaluated. A considerable reduction in mean percentage of number, natural morphology of sperm, sperm motility and viability and serum testosterone concentration besides DNA injury increment among mice treating with PHZ in comparison with control group were observed. However, in PHZ+EP group the above mentioned parameters were improved. This study showed that PHZ caused induction of toxicity on sperm parameters and reduction of testosterone as well as the increment of MDA level and EP as an antioxidant could reduce destructive effects of PHZ on sperm parameters, testosterone level and lipid peroxidation.

Isolation and identification of viral hemorrhagic septicemia virus (VHSV) from farmed rainbow trout (Oncorhynchus mykiss) in Iran.

Viral hemorrhagic septicemia virus (VHSV) is a rhabdovirus that causes one of the most important fish diseases in rainbow trout (Oncorhynchus mykiss) production industry. During the present study from October 2014 to July 2015, the virus causing viral hemorrhagic septicemia (VHS) was isolated and identified in rainbow trout farms from five of sixteen farms experiencing mass mortalities in six provinces of Iran with major trout production. Cumulative mortalities at VHSV-positive farms ranged from 30 to 70%. Clinical signs of disease included exophthalmia, petechial hemorrhages in the mandible and around the eyes, a swollen abdomen and darkening of the integument, widespread petechiae of the musculature and pyloric regions, severe congestion of the kidney, and pale enlarged livers. In addition, histopathologic examinations of tissues showed severe lesions in muscle, kidney and liver, which were compatible with those already described for VHS. Furthermore, homogenates tissues of diseased fish induced cytopathic effects (CPE) in CHSE-214 cells, and confirmatory diagnosis of VHS was made by RT-PCR reactions. To our knowledge, this is the first report of isolation and identification of VHSV from farmed trout in Iran, which may have originated from Europe.

Molecular identification and phylogenetic analysis of Lactobacillus and Bifidobacterium spp. isolated from gut of honeybees (Apis mellifera) from West Azerbaijan, Iran.

Polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) and phylogenetic analysis were used for molecular identification of lactic acid bacteria (LABs) isolated from Apis mellifera. Eighteen honeybee workers were collected from three different apiaries in West Azerbaijan. LABs from the gut of honeybees were isolated and cultured using routine biochemical procedures. Genomic DNA was extracted from LABs and a fragment of 1540 bp in size of 16S rRNA gene was amplified. PCR products were digested using HinfI endonuclease and digested products with different RFLP patterns were subjected to nucleotide sequencing and phylogenetic analysis. The results revealed that Lactobacillus and Bifidobacteria spp. are were the most abundant LABs in honeybee gut. Phylogenetic analysis showed that both Lactobacillus and Bifidobacterium were closely clustered with high similarity percentage with the same bacteria isolated from honeybees' gut elsewhere. It was concluded that LABs isolated from honeybees had low sequence divergence in comparison with LABs isolated from other sources such as dairy products.