Iranian journal of veterinary research最新文献

Background: Leptospira, a pathogenic bacterial organism, causes human leptospirosis when it infects mammals. There are different serovars of Leptospira, each of which is adapted to a specific animal host. Adaptation occurs dynamically in different hosts and changes the distribution and clinical manifestations of serovars. Variable-number tandem repeats (VNTRs) have been used to identify Leptospira interrogans strains.

Aims: The purpose of this study was to conduct seroepidemiology and molecular typing of pathogenic Leptospira isolated from domestic animals in Gilan province.

Methods: A total number of 250 urine and blood samples were collected from domestic animals (horse, bovine, and sheep). Urine samples were cultured in EMJH medium. Using the microscopic agglutination test (MAT) and the Multilocus VNTR Analysis (MLVA) methods, unknown samples were analyzed to determine Leptospira serovars.

Results: Based on our data, Leptospira Autumnalis was mainly found in horses and dominated at 50%, followed by Leptospira Canicola at 29.16%. MAT results showed that among 87 animals with the seropositive, the horses showed the highest level of contamination at 40%, and this difference was not significant (P=0.152).

Conclusion: According to our data, Leptospira Autumnalis was mainly found in horses, and the bacteria were shed in urine, suggesting that horses could transmit leptospirosis. The results of this study confirm that MLVA can replace time-consuming methods such as MAT because it is a simple and fast technique.

Background: Mammary tumors are the most common tumors in female dogs. An early diagnosis makes the treatment easier.

Aims: The present study aimed to assess heat shock protein D1 (HSPD1) expression in canine mammary tumors.

Methods: Canine mammary tumor (CMT) samples were collected from clinics after surgery. Expression of HSPD1 transcript in CMT and apparently healthy mammary tissues was analyzed by SYBR green based real-time PCR (qRT-PCR). Further, gene encoding the immuno-dominant region of HSPD1 was cloned using the expression vector pPROEX-HTa and expressed in a prokaryotic system and recombinant HSPD1 (rHSPD1) was purified by affinity chromatography. Hyperimmune serum was raised against rHSPD1 in mice, and immunohistochemistry was standardized to assess the expression of this protein in various histotypes of canine mammary tumors.

Results: An elevated HSPD1 mRNA expression (5.973 ± 0.862 folds) was observed in canine mammary tumors. Upon purification, a 60 kDa recombinant protein was obtained and confirmed by Western blotting. In 83.3% of healthy mammary tissues, a mild/feeble HSPD1 expression was observed whereas, a strong expression of HSPD1 was seen in 80% CMT samples.

Conclusion: The findings suggested that HSPD1 could be used as a molecular marker for canine mammary tumors.

Background: Uterine leiomyosarcoma is a malignant neoplasm arising from leiomyocytes in the uterus.

Case description: A 12-year-old mixed-breed intact female dog was presented with vaginal bleeding for the past 2 weeks. Abdominal ultrasonography revealed a mixed echogenic mass in the uterus, and right lateral abdominal radiography showed a large mid-abdominal mass. Exploratory celiotomy was performed, revealing a solid mass in the entire uterine body and both uterine horns. Ovariohysterectomy was performed and the uterine mass sample was microscopically examined.

Findings/treatment and outcome: The mass was composed of a dense proliferation of neoplastic cells that display two morphologic features; spindloid and epithelioid appearances. Additionally, chondroid-like matrix was noted. Both components were negative for cytokeratin AE1/AE3, and strongly positive for vimentin. The spindloid component was strongly positive for α-SMA, while the epithelioid-like component was moderately positive for alpha-smooth muscle actin. These findings are suggestive of a leiomyosarcoma. Although the recovery immediately following surgery was smooth and without complications, the dog's condition worsened over the following month, ultimately leading to euthanasia.

Conclusion: This report describes the histopathologic features and clinical outcome of an unusual variant of uterine leiomyosarcoma in a dog.

Background: To meet up the demand gap of milk in Bangladesh, short-term, midterm, and long-term goal have been set up by the government through crossing with Bangladeshi local cattle and high-producing foreign cattle like Friesian, Jersey, Sahiwal, etc.

Aims: The purpose of this study was to identify the single nucleotide polymorphisms (SNPs) in the FASN gene, to check the structural and functional impact of mutant proteins on milk production traits that are significantly associated.

Methods: Four SNPs were identified in exons 26, 36, 38, and 41 of the FASN gene using pooled DNA sequencing, but only one SNP g.17924 A>G was a non-synonymous that changed the amino acid threonine to alanine in the FASN protein and the other three SNPs were silent mutations. Structural and functional prediction analysis were done with a series of techniques to detect remote protein homology and predict structures, structural integrity, structure quality, protein stability, protein motion, flexibility, and stability impact, conservation profile and finally molecular dynamics simulations for wild-type and mutant protein expression differences.

Results: The non-synonymous g.17924 A>G mutation showed a clear difference between wild and mutant proteins, indicating the impact on the observed phenotype. Then, SNP g.17924 A>G was genotyped in 100 milking cows aiming to check the association effects. SNP g.17924 A>G was found to have significant allele substitution effects on milk yield traits.

Conclusion: Our results suggest that the identified polymorphism affects milk yield traits in the studied population and could be used as genetic marker for cattle selection processes aiming to increase productivity.

Background: Certain limitations are associated with all kinds of diagnostic techniques used for the identification of bovine cytological endometritis under field conditions. Leukocyte esterase (LE) tape of urinary test strips has been used to detect cytological endometritis with an acceptable level of agreement with endometrial cytology in lactating dairy cows; however, the sampling procedure of LE tape strips is still time-consuming and not advisable for use at the farm level.

Aims: The present study was designed to provide a simpler and faster LE test to improve the field applicability of LE tape strips for the detection of bovine cytological endometritis.

Methods: Endometrial samples were collected using cytobrush and modified LE tape techniques from slaughtered (n=74) and live dairy cows (n=43). The urinary strip's LE tape was affixed onto the cytobrush rod head using a hot glue adhesive. This modification created the possibility of rapid reading of the intensity of color change of LE tapes.

Results: Considering the cytology as the reference method, the sensitivity and specificity of the LE method for slaughterhouse samples were 82% and 73%, respectively. The agreement between endometrial cytology and the modified LE technique, considering the combination of slaughterhouse and clinical samples, was found to be moderate (=0.51; P<0.0001) in the present study.

Conclusion: A rapid leukocyte esterase test can be used as a cow side and potential alternative to endometrial cytology; however, the efficacy of this method should further be evaluated based on the conception rates in dairy cows.

Background: The impact of different physical and/or chemical treatments in cereal grains on starch morphology and ruminal digestibility has been evaluated.

Aims: The effect of chemical and/or physical treatments on starch and protein molecular appearance and the ex-vivo digestibility of barley grain was studied.

Methods: Treatments were: steam-flaked barley grain (SFB), SFB treated with ammonium bicarbonate (A), urea (U), and malic acid (M) (SFB_AUM), SFB treated with A, U, and lactic acid (L) (SFB_AUL), steam-infrared heated-flaked barley grain (SIFB), SIFB treated with A, U, and M (SIFB_AUM), and SIFB treated with A, U, and L (SIFB_AUL). Chemicals including A, U, M, and L were used as 56, 8, 10, and 10 g/kg dry matter (DM), respectively. Chemical composition and molecular morphology were determined using scanning electron microscopy and Fourier-transform infrared spectroscopy (FTIR). In situ mobile bag technique and in vitro batch culture procedure were used to estimate ruminal and post-ruminal digestibility.

Results: Crude protein (CP) and starch concentrations in SFB_AUL were higher than the others (P<0.05). Starch granule morphology and protein structure were altered in the chemically treated samples. The potentially digestible fraction of DM was the highest in the SFB_AUM (P<0.05). Ruminal disappearance of DM, CP, and starch was improved in SFB_AUL and SIFB_AUL compared with other groups (P<0.05). The highest post-ruminal digestibility of starch and CP was observed in SIFB_AUL and SIFB (P<0.05).

Conclusion: Present results indicate that chemical processing with L and applied steam-infrared heated-flaked in barley grain may improve in vitro digestibility of starch and CP and increase granule sizes.

Background: Domestic pigeons (Columba livia domestica) are the oldest domesticated birds worldwide, harboring many zoonotic parasites and posing potential public health threats.

Aims: To investigate cryptosporidiosis in domestic pigeons in Tabriz, Iran, 100 privately owned pigeons presenting weight loss and diarrhea were tested for Cryptosporidium spp. through parasitological, histopathological, and molecular tests.

Methods: Modified Ziehl-Neelsen-stained fecal smears and histological sections of the trachea and small intestine were examined microscopically. Genomic DNA of fecal and tracheal specimens was examined by nested conventional PCR targeting 18S rDNA, followed by Sanger sequencing of histopathology-confirmed samples and phylogenetic analyses.

Results: All pigeons were positive at PCR in their feces and trachea. Oocysts similar to the size of Cryptosporidium species were observed in stained fecal smears of 62% of pigeons. At the histopathological examination, Cryptosporidium-organisms were observed on the apical epithelial surfaces of the small intestine in 84% and trachea in 78% of pigeons. In 23 pigeons, simultaneous tracheal and intestinal cryptosporidiosis was determined. The lesions in affected tracheas and small intestines included hyperemia, villous atrophy and fusion, dilatation of intestinal crypts, irregular epithelial hyperplasia, and sloughing. Diffused mixed inflammatory cell infiltration in the lamina propria was observed, with dominant lymphocytes, plasma cells, and lower numbers of heterophils. Consensus sequences of detected parasites revealed infection with Cryptosporidium parvum and Cryptosporidium meleagridis.

Conclusion: Considering the high frequency of cryptosporidiosis reported here in symptomatic birds and that both identified Cryptosporidium species are zoonotic parasites, findings claim a public health risk assessment of this species of animals.

Background: Staphylococcus aureus, one of the causes of food poisoning, plays an important role in causing gastrointestinal inflammation.

Aims: Given the spread of antibiotic resistance in S. aureus, the present study aimed to investigate the effect of rosmarinic acid (RA) loaded magnetic nanoparticles (Fe₃O₄NPs@RA) on inhibiting the growth and biofilm formation of S. aureus isolated from meat samples.

Methods: Fe₃O₄NPs@RA have been synthesized and their antimicrobial activities were investigated against S. aureus isolated from poultry meat by broth micro-dilution. The anti-biofilm effect of these nanoparticles and their effect on the expression level of biofilm-associated genes were investigated using microplate and real-time PCR methods. The killing properties of Fe₃O₄NPs@RA against test bacteria investigated by time-kill assay.

Results: The minimum inhibitory concentration (MIC) of Fe₃O₄NPs@RA against S. aureus isolates ranged from 31.2-125 µg/ml. Also, the treatment with a sub-MIC concentration of Fe₃O₄NPs@RA prevented the formation of biofilm by 50-82%, in different isolates and downregulated the expression level of icaA and icaD. Also, the treatment with the MIC concentration of Fe₃O₄NPs@RA caused a 2.4-fold decrease in the population of living bacteria after 4 h and the number of living bacteria decreased more than 99% after 8 h. In the cytotoxicity assay, during 48 h, Fe₃O₄NPs@RA had no cytotoxic effect on HEK-293 cells at concentrations lower than of 300 µg/ml.

Conclusion: The results of the present study showed that Fe₃O₄NPs@RA were effective in inhibiting the growth and biofilm formation of S. aureus isolates and could be further investigated as an option for controlling S. aureus in food samples.

Background: Leishmania spp. are the cause of a common zoonotic illness. Dogs are the main reservoirs of the parasites, which play a considerable role in infecting humans and other hosts.

Case description: A 2-year-old male dog with evident acute skin lesions and ulcerative nodules on the face was referred to a small animal hospital in Mazandaran province, Iran. In order to detect Leishmania parasites, the popliteal lymph node (LN) was sampled for the microscopical examination and the PCR reaction. Also, the hematological and biochemical parameters were measured.

Findings/treatment and outcome: Light microscopy on the LN sample stained with Giemsa revealed Leishman bodies inside and outside of macrophages. Laboratory findings showed mild leukocytosis, lymphocytosis, neutrophilia, low hematocrit, hyperglobulinemia, hyperproteinemia, hypoalbuminemia, declined albumin/globulin ratio, and hyperglycemia. The PCR and sequencing results confirmed that Leishmania infantum was the causative agent.

Conclusion: Cutaneous leishmaniasis due to L. tropica and L. major is prevalent in humans and dogs in Iran. In this report, a generalized skin disease was evident caused by L. infantum, a viscerotropic parasite species. In addition to cutaneous signs, leukocytosis and anemia concordance with the change in biochemical parameters supported a visceral invasion. Therefore, this report is significant as the cutaneous form of the disease may also imply a complicated visceral illness. In Mazandaran province, the visceral type has been rarely reported. As a result, this form of leishmaniasis in dogs raises concerns about the possibility of zoonotic transmission and may threaten public health.

Background: Honeybee (Apis mellifera) venom contains several biologically active peptides, including melittin, apamin, enzymes, and non-peptide components. Due to its biologically active components, honeybee venom demonstrates antibacterial, antiviral, anti-inflammatory, and cytotoxic properties.

Aims: This study aimed to investigate the cytotoxic effects of honeybee venoms collected from different provinces in Turkey on colorectal adenocarcinoma (Caco-2) and glioblastoma multiforme (T98G) cell lines.

Methods: The apamin, phospholipase A2, and melittin contents of honeybee venoms were analyzed using a high-pressure liquid chromatography (HPLC) variable wavelength detector. The viability of cells after 24-hour exposure to honeybee venoms was determined using the 3-(4,5-dimethylthiazol-2-yl) (ΜTT), neutral red (NR), and dehydrogenase leakage (LDH) assays.

Results: Content analysis showed that Mugla had the highest apamin content, while Denizli had the highest phospholipase A2 and melittin contents among the analyzed bee venoms. For Caco-2 cells, the lowest inhibitory concentration (IC₅₀) values were observed in the Denizli venom group, with MTT and LDH results of 12.42 ± 0.19 µg/ml and 8.19 ± 0.61 µg/ml, respectively. For T98G cells, these values were 5.98 ± 0.40 µg/ml and 5.04 ± 0.17 µg/ml, respectively.

Conclusion: These findings indicate that honeybee venoms from different provinces contain varying levels of apamin, phospholipase A2, and melittin. The cytotoxic effects observed on Caco-2 and T98G cell lines suggest that honeybee venom may have potential as an anticancer agent.