The first molecular isolation of Halicephalobus gingivalis from horses in Iran.

BACKGROUND: Parasitic infections pose significant threats to humans' and animals' well-being worldwide. Among these parasites, Halicephalobus spp., a genus of nematodes, has gained attention due to its ability to cause severe infections in various animal species, including horses. OBJECTIVE: This study aimed to determine the prevalence of Halicephalobus spp., specifically focusing on Halicephalobus gingivalis in horses. MATERIALS AND METHODS: In July 2022, a cross-sectional study was conducted in northern Iran to determine the prevalence of Halicephalobus spp. Using standard coprological techniques, 141 fecal samples from randomly selected horses were analyzed for GI helminth eggs. The Halicephalobus spp. eggs present in faeces were identified by molecular methods. Polymerase Chain Reaction (PCR) was used to amplify the partial 5' variable region (~ 390 base pairs) of 18 S DNA using SSUA_F and SSU22_R primers. Furthermore, the PCR products obtained were sequenced, and phylogenetic analysis was performed using available sequences from GenBank. RESULTS: Microscopic examination of 141 fresh faecal samples revealed 5 fecal samples were infected with small ellipsoidal nematode eggs ranging between 40 and 50 × 50-60 µm. This study's PCR amplicons showed ~ 390 bp bands on 2.0% agarose gel. A partial sequence of 18 S DNA (363 bp) was obtained herein (GenBank accession no. OQ843456). CONCLUSION: Overall, using molecular tools represents a significant step forward in diagnosing and managing the Halicephalobus gingivalis infections in horses.

Detection of Hepatozoon spp. in dogs in Shiraz, southern Iran and its effects on the hematological alterations.

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

Molecular and hematological investigation of Trypanosoma evansi infection in Iranian one-humped camels (Camelus dromedarius).

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

Polymorphism in neutrophil cytosolic factor 4 (NCF4) of dairy cows had mastitis in previous lactations, and the relationship with the respiratory burst.

Nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase), as a key factor in innate immunity, consists of several components, one of them is p40phox which is encoded by neutrophil cytosolic factor 4 (NCF4). Respiratory burst and reactive oxygen species (ROS) production are antimicrobial mechanisms associated with NADPH oxidase. This study evaluated the effects of g.18174 A > G and g.18270C > T single-nucleotide polymorphisms (SNP) in NCF4 on bovine mastitis and the respiratory burst capacity of neutrophils. SNPs of 160 dairy cattle were determined using a novel PCR-RFLP protocol by employing restriction enzymes, MboI and FokI. Also, the flow cytometry measured respiratory burst in 82 blood samples. Our results indicated that only g.18174 A > G SNP reduced the respiratory burst capacity. However, both SNPs were not significantly correlated with clinical mastitis. We concluded that g.18174 A > G decreases the function of NADPH oxidase. However, both SNPs were not significantly correlated with clinical mastitis.

Theileria equi in the horses of Iran: Molecular detection, genetic diversity, and hematological findings.

In all equids worldwide, Theileria equi and Babesia caballi are believed to be two important erythrocytic protozoa that cause equine piroplasmosis. In addition, it was recently discovered that Theileria haneyi is another potential equine piroplasmosis (EP) agent. Ixodid ticks are the major vectors of these parasites. Equine piroplasmosis is of international importance and affects enormously the equine industry. In this study, for the first time, molecular prevalence and genetic diversity of piroplasma parasites (T. equi and B. caballi) in horses from Fars province (south of Iran) were determined. Also, hematological alterations of naturally infected horses were analyzed. PCR positive horses showed anemia, thrombocytopenia, leukocytosis with a left shift of neutrophilia, and monocytosis. PCR results revealed that, from 133 blood samples of horses, 40 samples were positive (30.07%). The occurrence of T. equi in this area (30.07%) was more than the national average prevalence of T. equi (24.11%), but B. caballi prevalence in study area (0%) was less than the average of previous studies in Iran (5.47%). Our findings revealed that the T. equi was widespread in Fars province of Iran. PCR products of 18S rDNA and EMA-1 genes of T. equi strains were sequenced successfully. All 18S rDNA sequences collected in this experiment revealed 100% similarity together. According to the phylogenetic tree constructed using the 18S rDNA gene, Iranian T. equi is clustered with strains from Cuba (KY111762, KY111761) and USA (CP001669, JX177672). So, this could be concluded that T. equi studied in this research, and those strains are initiated from a common T. equi ancestor at an unknown time ago. Also, the phylogenetic tree based on EMA-1 gene demonstrated a genetically diverse population of Iranian T. equi strains (10 different genotypes). As EMA-1 is one of the most immunogenic antigens in this parasite, such variability could be a concern about the efficacy of T. equi vaccines. Finally, more studies on equine piroplasmosis in the provinces of the southern region of Iran are recommended to create a better vision of disease in this region.

First report of ocular histoplasmosis in a horse from Iran: molecular, clinical and pathological findings.

Histoplasma capsulatum is a dimorphic fungus that is traditionally classified in three varieties: Hc var. capsulatum, Hc var. duboisii, and Hc var. farciminosum (HCF). Cytology, hematology, pathology, polymerase chain reaction (PCR), sequencing, and phylogenetic analyses were applied on samples collected from the blood and the eye of a horse with pustular lesions and ocular discharge. Physical examination and cytopathological tests showed H. capsulatum infection. Additionally, the results of two PCR tests confirmed H. capsulatum infection. The phylogenetic tree of the internal transcribed spacer sequence of Iranian H. capsulatum showed homology with the HCF variety. For the first time, H. capsulatum infection in the eye of a horse from Iran was detected and phylogenetically analyzed. This study revealed that H. capsulatum could establish infection in Iranian animals in addition to people, and indicated the role of soil enriched with bird dropping in the preparation of a favorable environment for H. capsulatum propagation. Further investigations are required to clarify the natural history and risk factors associated with histoplasmosis in Iran.

Intrauterine infusion of blood serum of dromedary camel improves the uterine health and fertility in high producing dairy cows with subclinical endometritis.

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

A study on the oral and cloacal bacterial flora of Mugger crocodiles (Crocodylus palustris) in the Negour protected area, Iran.

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.

Protective effects of crocin on testicular torsion/detorsion in rats.

Oxidative stress, caused by extreme accumulation of un-scavenged reactive oxygen species, plays an integral role in the Ischemia-Reperfusion (I/R) injury to the testicles following testicular torsion. The current research aimed to examine the protective effects of crocin as a natural antioxidant on testicular I/R injury in rats. Animals were divided randomly into five groups (seven each): (1) sham group, (2) torsion/detorsion (T/D) group, (3) intact group with 100 mg/kg crocin, (4) and (5) T/D groups followed by treatment with two different doses of crocin (50 and 100 mg/kg (IP)). I/R injury was induced by 720° clockwise torsion of the left testicles for 2 h. After 24 h of reperfusion, blood samples and epididymal sperms were collected to measure biochemical (GPx, SOD, and MDA), hormonal (testosterone), and sperm parameters (total sperm recovery, motility, viability, and morphology). Moreover, affected testicles were subjected to histopathology examination. I/R injury caused a significant reduction in sperm characteristics (except for morphology) (P < 0.05), which could not be significantly improved by crocin administration at either dose (P > 0.05). Johnsen's testicular score, mean seminiferous tubular diameter, and germinal epithelial cell thickness were significantly decreased in the T/D group compared to the intact and sham groups. However, crocin could significantly improve the histopathological parameters in both treatment groups compared to the T/D group (P < 0.05). T/D reduced SOD and GPx activity and testosterone level significantly (except for GPx) compared to the sham group (P < 0.05). However, crocin administration could significantly reverse them. Also, crocin reduced the amount of MDA significantly in the high-dose treatment group in comparison to T/D group (P < 0.05). The results of the current study revealed that crocin could be a promising agent to protect against I/R injury following surgical correction of the testicular torsion.

Class 1 integron causes vulnerability to formaldehyde in Escherichia coli.

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

High incidence of multidrug resistance and class 1 and 2 integrons in Escherichia coli isolated from broiler chickens in South of Iran.

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

Molecular detection and phylogenetic analysis of 'Candidatus Bartonella merieuxii' in dogs and its effect on hematologic parameters.

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

The occurrence of hemotropic Mycoplasma ovis-like species in horses.

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

Molecular Characterization of Fasciola spp. from a Donkey (Equus asinus) Using Partial Sequencing of cox1 and nad1.

BACKGROUND: Fasciola hepatica as an important parasite affects health of humans and animals in some tropical and subtropical areas of the world, including Iran. Little is known about the molecular diversity of Fasciola in Equidae. Therefore, this study aimed to characterize the genetic polymorphisms among parasites. METHODS: Eight adult Fasciola spp. isolates were collected from a working donkey after necropsy in Shiraz, southwestern Iran, in 2018. Primarily, various parameters were measured morphologically. Subsequently, DNA was extracted from each fluke and molecular markers of cytochrome C oxidase (cox1) and NADH dehydrogenase 1(nad1) from individual Fasciola isolates were amplified using PCR assay and sequence data were employed for molecular and phylogenetic analysis. Genetic diversity between isolates was evaluated by comparing the sequences of these two mitochondrial regions. RESULTS: Based on the morphological and analyzed mitochondrial sequences, all of eight donkey isolates (100%) were identified as F. hepatica. Moreover, nine and five nucleotide polymorphisms were identified in the cox1and nad1 region sequences, respectively. CONCLUSION: Accordingly, phylogenetic data revealed five and four haplotypes among donkey isolates based on the cox1and nad1 markers. Similarly, some of these haplotypes have been previously reported from different host species in Iran as well as all around the world.

Antimicrobial Resistance and Incidence of Integrons in Aeromonas Species Isolated from Diseased Freshwater Animals and Water Samples in Iran.

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

'Candidatus Bartonella dromedarii' in the dromedary camels of Iran: Molecular investigation, phylogenetic analysis, hematological findings, and acute-phase proteins quantitation.

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

Effect of catecholamine stress hormones (dopamine and norepinephrine) on growth, swimming motility, biofilm formation and virulence factors of Yersinia ruckeri in vitro and an in vivo evaluation in rainbow trout.

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

First isolation and identification of Aeromonas veronii and Chryseobacterium joostei from reared sturgeons in Fars province, Iran.

The purpose of the present study was to isolate and identify the pathogenic agents in Acipenser stellatus (Pallas, 1771) and Huso huso, (Linnaeus, 1758) reared in the south of Fars province, Iran which have shown infectious disease signs. Samples from spleen and kidney of 32 fishes showing septicemia symptoms such as decreasing of appetite, unbalanced swimming, expanded wounds, and petechia on the body surfaces, pectoral fins rot, visceral hemorrhage, bleeding on the spleen, and heart ascites were collected. Then samples were cultured on brain heart infusion agar growth media, stain and biological and biochemical tests on purified bacteria were performed. On the other hand, 16S rDNA region of the isolated organism was amplified using PCR. The amplified gene fragment was sequenced and evolutionary history was inferred by phylogenetic tree construction using neighbor-joining method. Results indicated that two bacterial species including Chryseobacteriumjoostei which isolated from the kidney of stellate sturgeon (43.00%), and Aeromonasveronii which isolated from the spleen of both sturgeon species (75.00% and 31.00% from beluga and stellate sturgeon, respectively), were recognized. Phylogenetic tree analysis showed that Fars isolated organisms including A. veronii and C. joostei had highest similarity with A. veronii bv veronii and C. joostei isolated from France, respectively.

Effect of quorum quenching bacteria on growth, virulence factors and biofilm formation of Yersinia ruckeri in vitro and an in vivo evaluation of their probiotic effect in rainbow trout.

Five N-acyl homoserine lactone-degrading bacteria (quorum quenching (QQ) strains) were selected to evaluate their impacts on growth, virulence factors and biofilm formation in Yersinia ruckeri in vitro. No difference was observed among the growth pattern of Y. ruckeri in monoculture and coculture with the QQ strains. To investigate the regulation of virulence factors by quorum sensing in Y. ruckeri, cultures were supplemented with 3oxo-C8-HSL. The results indicated that swimming motility and biofilm formation are positively regulated by QS (p < 0.05), whereas caseinase, phospholipase and haemolysin productions are not influenced by 3oxo-C8-HSL (p > 0.05). The QQs were able to decrease swimming motility and biofilm formation in Y. ruckeri. QQ bacteria were supplemented to trout feed at 108  CFU/g (for 40 days). Their probiotic effect was verified by Y. ruckeri challenge either by immersion or injection in trout. All strains could significantly increase fish survival with Bacillus thuringiensis and Citrobacter gillenii showing the highest and lowest relative percentage survival (RPS) values (respectively, 85% and 38%). Besides, there was no difference between the RPS values by either immersion or injection challenge expect for B. thuringiensis. The putative involvement of the QQ capacity in the protection against Yersinia is discussed.

Characterization of polymorphism in the FSH receptor gene and its impact on some reproductive indices in dairy cows.

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