Lumpy jaw caused by Nocardia farcinica in a Holstein cow.

A 3-year-old Holstein cow was examined in an intensive system due to unilateral swelling in the mandible. A right mandibular mass was associated with painful mastication and Ptyalism. In palpation, the mass was raised, ulcerated, attached to the mandible bone and firm, approximately 17 × 12 × 10 cm3 in size. The lesion was sampled, and after routine bacteriology and histopathology procedures, the occurrence of lumpy jaw caused by Nocardia farcinica was confirmed. The bacterium was analysed using genome sequencing and new strain called Najm 114. Due to the risk of zoonosis of the isolated agent, the cow was euthanized. This is the first report of lumpy jaw caused by N. farcinica in a cow. This study showed that N. farcinica should be considered a possible etiological agent for lumpy jaw in cattle.

A rare case of Cysticercus tenuicollis infection in a neonate lamb: Evidence of prenatal transmission.

Cysticercosis develops in lambs following a Cysticercus tenuicollis infestation, which is the larval stage of Taenia hydatigena. A 7-day-old lamb was examined for depression, anorexia, fever (40.5°C), congested mucus membranes, reluctance to move, and a hunched back. Upon necropsy, congestion was noted in the intestines and brain, and the heart had a loose consistency. Soft and pulpy kidneys were evident coupled with watery intestinal contents. Epsilon toxin (Clostridium perfringens type D toxin) was detected using enzyme-linked immunosorbent assay. A transparent cystic structure was incidentally found attached to the pancreas, within which a scolex was well demonstrated upon histopathology. Chronic active peritonitis was diagnosed at the cyst attachment site. C. tenuicollis was confirmed by polymerase chain reaction and genome sequencing. This report describes prenatal transmission of C. tenuicollis in the present lamb, although this condition is quite rare.

Clinico-histopathological and phylogenetic analysis of protozoan epibiont Epistylis wuhanensis associated with crustacean parasite Lernaea cyprinacea from ornamental fish in Iran.

Parasitic infestations are one of the most concerning problems limiting ornamental fish farming. In addition to the direct economic losses due to the major mortalities, parasites may significantly negatively impact the body shape, coloration, growth rate, and health condition of the fish. The results of the study highlight the importance of comprehensive parasitological analysis in the diagnosis and treatment of parasitic infections in ornamental fish farms. The presence of multiple parasites in the affected fish emphasizes the need for effective biosecurity measures, such as extending the quarantine period for newly imported fish, closely monitoring fish populations, and implementing isolation units to prevent the spread of infections. By implementing these preventative measures, ornamental fish farmers can reduce the risk of parasitic infections and ensure the health and well-being of their fish populations. This, in turn, can lead to increased profitability and sustainability for their business. Overall, the current study aimed to conduct a clinical, histopathological, and phylogenetic analysis of the epibiont ciliated protozoan Epistylis wuhanensis and the copepod crustacean Lernaea cyprinacea in a freshwater ornamental fish farm in Iran. Furthermore, it provides valuable insights into the prevalence and impact of parasitic infections in ornamental fish farms and underscores the need for continued research and the development of effective preventative measures to address this issue. A total of 60 symptomatic freshwater ornamental fish, including 30 guppy (Poecilia reticulata) and 30 sailfin molly (Poecilia latipinna), were packed in polyethylene bags filled with oxygenated pond water and transported to the Ornamental Fish Clinic, Faculty of Veterinary Medicine, University of Tehran, for parasitological analysis. Following the clinical examination, histopathological analysis was performed on 10% NBF (neutral buffered formalin)-fixed samples from affected tissues, including the skin, skeletal muscle, and liver, to identify any pathological changes associated with the parasitic infections. Furthermore, the DNA was extracted from the 99% ethanol-fixed samples using a commercial DNA extraction tissue kit (SinaPure DNA, Iran), and PCR was performed using Peri18S-F1 (5'-ACC TGG TTG ATC CTG CCA GT-3') and Peri18S-R1 (5'-TGC AGG TTC ACC TAC GGA AA-3') (first reaction), and Peri18S-F2 (5'-CCG CGG TAA TTC CAG CTC-3') and Peri18S-R2 (5'-GAT CCC CTA ACT TTC GTT CTT GA-3') (second round) primers for the identified parasites. Finally, the PCR products were sequenced using Sanger dideoxy sequencing methods, and the resulting sequences were compared to sequences in the BLAST search program to provide a comprehensive picture of the current parasite-based disorder. The crustacean L. cyprinacea and the epibiont sessilid E. wuhanensis were identified in the examined ornamental guppy (6/30) and sailfin molly (6/30), with an overall parasitic prevalence of 20.00% (12/60). Ciliates were found in all tissue lesions but not in fish without lesions. A great number of the ciliated protozoan E. wuhanensis were found attached to the integumentary area of L. cyprinacea. Microscopically, oval to round granulomatous lesions were observed in cutaneous and skeletal muscles. Lymphoplasmacytic dermatitis and myositis were also observed. The crustacean L. cyprinacea serves as a mechanical vector for E. wuhanensis infection and spreads the disease in ornamental fish farming operations. For the first time in Iran, we successfully presented diagnostic morphological and molecular data on sessilids isolated from L. cyprinacea. Based on the findings of the current study, such parasitic infections may cause significant economic losses following invasion of the integument area of fish, eventually leading to death, if treatment is neglected or inadequate. Furthermore, the findings of the analysis were used to develop effective diagnostic approaches for the affected fish, as well as recommendations for improved health conditions to prevent future outbreaks of parasitic infections. However, further research is needed to determine the precise mechanisms of crustacean attachment and host-crustacean-peritrich protozoan interactions. Furthermore, the direct and indirect effects of various environmental factors on the emergence and spread of the current disease should be considered.

Acute cysticercosis caused by Cysticercus tenuicollis in lambs.

Twelve 30- to 75-day-old mixed breed lambs were examined in an intensive system because of sudden recumbency and death. Clinical examination revealed sudden recumbency, visceral pain, and auscultation of respiratory crackles. Lambs died shortly (between 30 minutes and 3 hours) after the onset of clinical signs. The lambs were necropsied, and after routine parasitology, bacteriology and histopathology procedures, the occurrence of acute cysticercosis caused by Cysticercus tenuicollis was confirmed. The use of the suspect infested feed (newly purchased starter concentrate) was discontinued and other lambs of the flock were treated with praziquantel (15 mg/kg, single dose, orally). After these actions, no new cases were observed. The present study showed the importance of preventive measures against cysticercosis in intensive sheep farming systems which include proper storage of feed, preventing feed and environmental access by potential definitive hosts, and implementing consistent parasite control programs in dogs that are in contact with sheep.

Construction of live-attenuated Trueperella pyogenes by antibiotic treatment and sequential passage: methods for vaccine development.

Trueperella pyogenes (T. pyogenes) is a zoonotic pathogen that is cause a variety of pyogenic diseases in animals. The complex pathogenicity and various virulence factors are important challenges to produce an effective vaccine. According to previous trials, inactivated whole-cell bacteria or recombinant vaccines were unsuccessful in preventing disease. Thus, this study aims to introduce a new vaccine candidate based on a live-attenuated platform. For this purpose, first T. pyogenes was subjected to sequential passage (SP) and antibiotic treatment (AT) to lose their pathogenicity. Second, Plo and fimA expressions as virulence genes were evaluated by qPCR and then mice were challenged with bacteria from SP and AT culture by intraperitoneal route. Compared to the control group (T. pyogenes-wild type), plo and fimA gene expressions were downregulated and vaccinated mice have a normal spleen appearance in contrast to the control group. In addition, there was no significant difference between bacterial count from spleen, liver, heart and peritoneal fluid in vaccinated mice and the control group. In conclusion, this study introduces a new T. pyogenes vaccine candidate based on a live-attenuated strategy that mimics natural infection without pathogenicity for further investigation on vaccines against T. pyogenes infections.

A quantitative prevalence of Escherichia coliO157 in different food samples using real-time qPCR method.

Escherichia coli serogroup O157 is the main causative agent of several intestinal and extra-intestinal foodborne diseases in humans through consumption of low-dose contaminated foods such as milk, beef, and vegetables. To date, studies regarding the quantitative prevalence of E. coli O157 in foods are so limited. Therefore, this study aimed to evaluate the quantitative prevalence rate of E. coli serogroup O157 in raw milk (n = 144), vegetable salad (n = 174), and minced beef samples (n = 108) using the real-time qPCR SYBR green melting curve method targeting the rfbA gene. First, we evaluated the method and found a sensitive and specific qPCR assay with 1 log of CFU/ml detection limit to detect E. coli O157 (Tm = 80.3 ± 0.1°C). About 2.77%, 10.18%, and 9.19% of raw milk, minced beef, and vegetable salad samples, respectively, were contaminated with E. coli O157. Minced beef and vegetable salad samples were significantly more contaminated than raw milk samples. Population average of E. coli O157 in raw milk, minced beef, and vegetable salad samples were 2.22 ± 0.57, 3.30 ± 0.40, and 1.65 ± 0.44 log CFU/ml or gr, respectively. Significantly higher levels of population of E. coli O157 were observed in minced beef samples. Minced beef can be regarded as the main food in the transmission of this foodborne pathogen. Routine quantitative rapid monitoring is strongly suggested to be carried out to prevent foodborne diseases caused by E. coli O157.

Isolated Bacteria from the Uteri of Camels with Different Reproductive Backgrounds: A Study on Sampling Methodology, Prevalence, and Clinical Significance.

The objectives of this study were to comparatively identify the common bacterial isolates from the uteri of camels coming from different reproductive backgrounds after standardizing the sampling method and to investigate the association of clinically measurable parameters with uterine colonization by these isolates. The uterine samples from 856 dromedary camels yielded a total of 17 different bacterial species with a higher proportion of sub-fertile camel uteri being colonized by bacteria (66.6%) as compared to nulliparous, recently calved, and those with unknown reproductive history combined (44.2%; p < 0.05). Camels with body condition scoring < 3 and those with a consistently echogenic appearance of the uterine lumen by sonography were more likely to be positive on uterine culture, while the presence of pus in uterine discharge was not associated with the odds of bacterial isolation (p > 0.05). While certain strains were more likely to be obtained from the uteri of the sub-fertile group (p < 0.05), embryo transfer to camels with a positive uterine culture in the absence of other gross reproductive pathologies did not necessarily affect the overall pregnancy rate compared to recipients with a negative uterine culture (p > 0.05). In conclusion, a relatively high bacterial load can be identified from the uteri of both sub-fertile and normal dromedary camels, with a higher frequency among the former. The uterine ultrasonography and evaluation of the body condition score can help in identifying the camels in which uterus is contaminated by bacteria.

Complete genome sequence of Himetobi P strain Sh.Moghaddam, isolated from the Laodelphax striatellus (small brown planthopper).

OBJECTIVE: Himetobi P virus (HiPV) is an insect virus belonging to the genus Cripavirus in the Dicistroviridae family within the Picornavirales order. Himetobi P strain. Sh.Moghaddam is the first study reported, was isolated from the Laodelphax striatellus (small brown planthopper) of an internal chicken organ in Iran. DATA DESCRIPTION: Genomic analysis showed a nucleotide identity of 93.16% with the family Dicistroviridae, genus Triatovirus, and species Himetobi P. The genome assembly comprised 9227 bp, with a 38.8% GC content. Annotation of the genome showed 2 ORF, a total of 2 genes: including 2 coding sequences (CDs) (total) and 8 Miss features. Thus, the whole-genome sequence presented in this study serves as a platform for detecting new genes that may contribute to the pathogenicity of the Himetobi P strain. Sh.Moghaddam.

Investigation of antimicrobial susceptibility and virulence factor genes in Trueperella pyogenes isolated from clinical mastitis cases of dairy cows.

Trueperella pyogenes is an opportunistic pathogen causing important diseases including mastitis and metritis in domestic animals such as dairy cows leading to prominent economic losses in food production industry. The aim of this study was to investigate bacterial species, antimicrobial susceptibility, and presence of virulence factor genes and genotyping of T. pyogenes isolates associated with summer mastitis cases from 22 different farms around Tehran, Iran. Fifty-five percent of dairy cows with clinical mastitis symptoms was infected by T. pyogenesis indicated that this pathogen is the most important contributor to clinical mastitis in dairy cows in the present study. A significant correlation was illustrated between presence of virulence factor genes of isolated pathogen, biochemical patterns, and the utter infected types. Multidrug resistance susceptibility observed between isolates indicated the important need for prudent use of antimicrobials in treatment of mastitis caused by T. pyogenes and increased concerning of consumer health associated with recent problems of antimicrobial resistance. The categorization of isolates was implemented into seven different clonal related types by COX-PCR at 80% of similarity cutoff with significance relationship to clonal types, CAMP test result and sampling time and biochemical profile. Regarding to the results obtained at the present study, T. pyogenes can be considered as an important typically cause of purulent and acute form of clinical bovine mastitis and loss of dairy productivity. Further studies with more sample size and high-throughput omic methods in various sampling time and areas are suggested for study of this pathogen precisely.

Complete genome sequence of Trueperella pyogenes strain Arash114, isolated from the uterus of a water buffalo (Bubalus bubalis) in Iran.

OBJECTIVE: Trueperella pyogenes has been considered a major causative agent of metritis, abortion, and death in a broad range of domestic and wild animals, including cattle, swine, sheep, goats, camels, buffalo, deer, antelopes, reptiles, and birds. DATA DESCRIPTION: Here, we report the complete chromosome sequence of Trueperella pyogenes strain Arash114, isolated from the uterus of a water buffalo (Bubalus bubalis) died due to the infection caused by this pathogen. The genome assembly comprised 2,338,282 bp, with a 59.5% GC content. Annotation of the genome showed 46 tRNA genes, 6 rRNA, 1 CRISPR and 2059 coding sequences. Also, several genes coding for antimicrobial resistance such as tetW and virulence factor including plo, nanH, nanP, cbp and 4 fimbrial proteins were found. This study will advance our knowledge regarding the metabolism, virulence factors, antibiotic resistance and evolution of Arash114 strain and serve as an appropriate template for future researches.

Genetic characterisation of methicillin-resistant Staphylococcus aureus and Staphylococcus pseudintermedius in pets and veterinary personnel in Iran: new insights into emerging methicillin-resistant S. pseudintermedius (MRSP).

OBJECTIVES: Methicillin-resistant staphylococci, including methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus pseudintermedius (MRSP), pose a threat to animal and human health worldwide. Veterinary staff and pets may play a role in the spread of resistant clones. METHODS: A total of 125 samples from veterinary staff (n=50), dogs (n=49) and cats (n=26) were investigated. Obtained isolates were tested for the methicillin resistance gene mecA and were subjected to multiplex PCR to differentiate coagulase-positive species. Following SCCmec and spa typing, isolates were tested for the presence of various toxin and virulence genes and phenotypic resistance to common antimicrobials. RESULTS: Overall, 4 MRSA were isolated from two veterinarians and two dogs and 19 MRSP were found in eleven dogs (12 isolates) and five cats (7 isolates). The MRSA isolates possessed sea (2) and eta (3) virulence genes and the MRSP isolates possessed sea (6), expA (15), expB (1) and siet (19) genes. SCCmec type II and three spa types (t186, t1816 and t10897) were identified in the MRSA isolates. Most of the MRSP isolates belonged to SCCmec types II (2 isolates) and V (10 isolates); however, the remaining 7 isolates were untypeable and contained class C1 mec. The majority of isolates were multidrug-resistant (MDR). CONCLUSION: These findings show that pets and veterinarians could be potential sources of MDR-MRSA and MDR-MRSP in Iran. Taken together, these findings warrant future investigations on the epidemiology and public-health significance of MDR-MRSA and MDR-MRSP both in veterinarians and companion animals in Iran.

New Brucella abortus S19 Mutant to Improve Distinction Between Infected and Vaccinated Animals.

BACKGROUND: Using Brucella abortus Strain 19 (S19) to control bovine brucellosis is restricted due to induce antibodies to the O-side chain of the smooth lipopolysaccharide (LPS) which may be difficult to differentiate vaccinated and infected animals. Furthermore, it is virulent for humans and can induce abortion to cattle. OBJECTIVES: The aim of this study was to employ gene knockout B. abortus S19 for the first time to eliminate diagnostic defects and obtain the attenuated mutant strain. MATERIAL AND METHODS: The wbkA gene, which is one of the LPS O-chain coding genes, was knocked out in vaccinal Brucella abortus S19. The proliferative response and immunoglobulin M production were analyzed in wbkA deletion strain-infected BALB/c mice. RESULTS: The loss of wbkA gene function resulted in induction of the splenocyte proliferative response in mice infected by the mutant S19 strain compare to those induced by parental S19 and RB51 strains. Moreover, wbkA mutant did not induce any IgM antibody response using the enzyme-linked immunosorbent assay. CONCLUSIONS: As a result, the new mutant S19 strain had deficiency in its LPS O-chain structure, besides cannot induce IgM response then, reduce mistakes to discriminate between vaccinated and infected animal, and also can be considered as a new vaccine candidate.

Genomic characterisation, detection of genes encoding virulence factors and evaluation of antibiotic resistance of Trueperella pyogenes isolated from cattle with clinical metritis.

Trueperella pyogenes is one of the most important microorganisms causing metritis in post-partum cattle. Co-infection with other bacterial species such as Escherichia coli or Fusobacterium necrofurom increases the severity of the disease and the persistence of bacteria in utero. The aim of this study was to investigate the frequency of T. pyogenes strains, and their virulence and antimicrobial resistant profiles in metritis cases. The study was carried out on 200 samples obtained from metritis discharges of postpartum cattle on 18 farms around Tehran, Iran. Sixty-five T. pyogenes isolates (32.5%) were identified, of which 16 isolates were detected as pure cultures and the other 49 isolates from cultures most commonly mixed with E. coli or F. necrofurom. In terms of diversity in biochemical characteristic of T. pyogenes strains, 8 different biotypes were identified among the isolates. Single or multi antimicrobial resistance was observed in 48 isolates (73.9%), which was mostly against trimethoprim sulfamethoxazole, azithromycin, erythromycin and streptomycin. The tetracycline resistance gene tetW and macrolide resistance genes ermB and ermX were detected in 30, 18 and 25 isolates, respectively. In the screening of genes encoding virulence factors, fimA and plo genes were identified in all tested isolates. Genes encoding nanP, nanH, fimC, fimG, fimE and cbpA were detected in 50, 54, 45, 40, 50 and 37 of isolates, respectively. Thirteen different genotypes were observed in these T. pyogenes isolates. A significant association between clonal types and virulence factor genes, biochemical profile, CAMP test result, severity of the disease and sampling time was detected.

Comparative study of Microsporum canis isolates by DNA fingerprinting.

Microsporum canis is a zoophilic fungus and it is an important agent of dermatophytosis. Cats act as important reservoirs. Clinically, it is too difficult to differentiate dermatophytosis caused by various species, also this fungus loses its morphological characteristics easily because of subculture; so using of rapid and accurate laboratory techniques for identifying the dermatophytes is important, therefore, RAPD-PCR was applied for the differentiation of the isolates. In this study, 10 M. canis isolates were detected in cats, dog, human, fox and rabbit at the Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran. For running the RAPD-PCR, PCR set system and three random primers OPU 15, OPU 13 and OPA 04 were used. Then phylogenetic tree and similarity coefficient table were drawn. The results showed that there were some common bands between M. canis isolates. There were some specific bands for each isolates, as well. Our study showed, despite the typical morphology of the whole isolates, they were placed in different branches in molecular typing.