Microbiology
Masoud Amini; Abdulghaffar Ownagh; amir Tokmachi; Manochehr Allymehr
Articles in Press, Accepted Manuscript, Available Online from 02 December 2023
Abstract
The poultry products are known as a resource of zoonotic and multi drug resistant pathogens, especially Salmonella spp. Using bacteriophages as an alternative antimicrobial agent are objective. Salmonella typhimurium isolate from turkey poults. The antibiotic susceptibility test was used identified the ...
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The poultry products are known as a resource of zoonotic and multi drug resistant pathogens, especially Salmonella spp. Using bacteriophages as an alternative antimicrobial agent are objective. Salmonella typhimurium isolate from turkey poults. The antibiotic susceptibility test was used identified the antibiotic resistance pattern of the isolates. The bacteriophage was purified, enhanced, and titrated using the Spot test and Double Layer Agar techniques after being isolated from a chicken slaughterhouse and sewage treatment facility. By determining the morphological characteristics of resulting plaque, the specificity and host range of the phage were studied on S. typhimurium isolates. The number of 22 suspected Salmonella isolates was confirmed biochemically positive in sample by cultures method. Nine of these isolates (40.90%) were identified as Salmonella typhimurium by Polymerase Chain Reaction. All of isolates (100%) were resistant to Chloramphenicol, Doxycycline, Kanamycin, Florfenicol, Rifampin, and Erythromycin. Seven isolates (77.70%) were resistant to Amoxicillin and Nalidixic acid. The plaques were present with a diameter of 3 ± 0.22 mm on the culture of 6 out of 9 (66.60%) isolates of S. typhimurium on the B.H.I broth medium using Double Layer Agar method. The amount of phage titer was 7.6 × 107 pfu/ mL and its Multiplicity of infection value was calculated as 5.06 × 10-2 based on obtained results. In place of antibiotics, the MDR S. typhimurium was successfully destroyed by the isolated bacteriophage from wastewater. In vitro settings were used in this investigation to identify the efficient bacteriophages against MDR S. typhimurium.
Microbiology
Hassan Tizfahm Tikmehdash; Alireza Dehnad; Nader Mosavari; Behroz Naghili Hokmabadi; Sanaz Mahmazi
Articles in Press, Accepted Manuscript, Available Online from 13 March 2024
Abstract
Glanders caused by Burkholderia mallei is one of the most dangerous zoonotic diseases in solipeds. Clinical diagnosis of this disease in its early stages in horses, is difficult. This study investigated serological and molecular identification of B. mallei in East Azerbaijan province. In the third and ...
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Glanders caused by Burkholderia mallei is one of the most dangerous zoonotic diseases in solipeds. Clinical diagnosis of this disease in its early stages in horses, is difficult. This study investigated serological and molecular identification of B. mallei in East Azerbaijan province. In the third and fourth quarters of 2020, throughout 2021, and in the first and second quarters of 2022, the complement fixation test (CFT) was performed on 350 horses. The malleination was used to confirm the positive CFT cases. Blood samples were taken for culture and for preparing serums to perform the ELISA. Deep eye discharge, nostril, cutaneous ulcers, and lymph fluid swabs were cultured, and finally, PCR was carried out. Eleven (3.14%) horses were CFT-positive. Based on the malleination on these 11 horses, six (1.7%) were affected by glanders, five (1.42%) were not affected (false positive), and one horse (0.28%) was CFT-negative despite exhibiting clinical signs. It was positive by malleination, ELISA, and PCR. A total of seven positive cases of glanders were diagnosed. B. mallei could not be isolated, but the B. cepacia complex was isolated in one case. Except for three cases (14.28%), the results of the CFT, mallein, and ELISA tests were consistent (71.42%). The amount of confidence interval (CI) was 95%. It is suggested that ELISA be used as a complement to CFT in screening and, if positive results are observed in one of the tests, the entire herd be examined more accurately using the mallein and western blot confirmatory tests.
Microbiology
saeed alamian; Karim Amiry; Afshar Etemadi; Maryam Dadar
Articles in Press, Accepted Manuscript, Available Online from 13 March 2024
Abstract
Bovine brucellosis, an infectious disease transmitted by Brucella melitensis and Brucella abortus, presents a significant risk as a zoonotic hazard that may result in adverse consequences for agricultural economics and animal health. The primary objective of this study is to present a comprehensive understanding ...
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Bovine brucellosis, an infectious disease transmitted by Brucella melitensis and Brucella abortus, presents a significant risk as a zoonotic hazard that may result in adverse consequences for agricultural economics and animal health. The primary objective of this study is to present a comprehensive understanding of the prevalence and features of Brucella strains within the industrial dairy farming sector in Iran. RBPT, SAT, and I-ELISA tests were used to confirm all seropositive animals. A total of 1311 bovine samples from seropositive animals (lymph nodes, milk, aborted fetuses, and abomasum contents) were collected from 224 farms in different provinces of Iran and examined bacteriologically. The discovered Brucella isolates were phenotyped and molecularly characterized. The isolates were all B. abortus or B. melitensis. Bacteria analysis revealed that 70.5% of seropositive farms tested positive for Brucella strains, predominantly B. melitensis biovar 1 (43.5%) and B. abortus biovar 3 (27.2%). Geographical distribution revealed that B. melitensis biovar 1 was the most common in dairy cow farms (16 provinces), followed by B. abortus biovar 3 (6 provinces). Furthermore, the prevalence of B. melitensis biovar 2, B. melitensis biovar 3, B. abortus biovar 1, B. abortus biovar 2 and RB51 vaccine were restricted to certain provinces. AMOS-PCR and Bruce-ladder PCR confirmed species identification. These results highlight the complexity of bovine brucellosis in Iran and illustrate that B. melitensis spreads from small ruminants to cattle. This study provides important epidemiological insights for targeting future brucellosis control programs in the Iranian dairy farms.
Microbiology
Leila Maleki; Amir Tukmechi
Volume 15, Issue 3 , March 2024, , Pages 159-164
Abstract
Staphylococcus aureus is an important pathogen causing a wide range of diseases in both humans and animals. The aim of this research was to screen the vancomycin resistance-associated genes in methicillin-resistant Staphylococcus aureus (MRSA) isolates from animals. A total of 400 nasal swab samples ...
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Staphylococcus aureus is an important pathogen causing a wide range of diseases in both humans and animals. The aim of this research was to screen the vancomycin resistance-associated genes in methicillin-resistant Staphylococcus aureus (MRSA) isolates from animals. A total of 400 nasal swab samples were collected from cattle, goats and sheep between February and August 2022 from both industrial and traditional livestock farms in West Azerbaijan province, Iran. Then, nasal swabs were cultured on mannitol salt agar and molecular analysis was performed after bacteriological examination to confirm the presence of S. aureus. The MecA gene was used to detect MRSA isolates, and two important vancomycin resistance-associated genes, namely vanA and vanB, were searched in the isolates. Out of 400 nasal swabs, 69 samples had S. aureus; of which seven isolates were resistant against methicillin. No vancomycin resistance-associated genes were detected in the MRSA isolates. Based on these findings, vancomycin could be used to treat infections caused by this bacterium.
Microbiology
Bhavinkumar Pankajbhai Katira; Bhaveshkumar Ishwarbhai Prajapati; Ratn Deep Singh; Sandipkumar Sureshbhai Patel; Kirankumar Motiji Solanki
Volume 15, Issue 2 , February 2024, , Pages 57-64
Abstract
Mastitis associated Klebsiella pneumoniae species were isolated from bovine milk to characterize virulence genes (wabG and kfuBC) and extended spectrum β-lactamase (ESBL) genes (blaCTX-M-1, blaCTX-M-2, blaCTX-M-9, blaTEM, blaSHV and blaOXA). A total number of 325 bovine milk samples (195 raw and ...
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Mastitis associated Klebsiella pneumoniae species were isolated from bovine milk to characterize virulence genes (wabG and kfuBC) and extended spectrum β-lactamase (ESBL) genes (blaCTX-M-1, blaCTX-M-2, blaCTX-M-9, blaTEM, blaSHV and blaOXA). A total number of 325 bovine milk samples (195 raw and 130 mastitic milk specimens) collected from Banaskantha, a milk-shed district of Gujarat, India, were included in the study. A total number of 27 K. pneumoniae isolates were recovered, consisting of 17 (62.96%) isolates from raw milk and 10 (37.03%) isolates from mastitic milk samples, giving an overall prevalence of 8.31%. Antibiotic sensitivity patterns revealed that 20 out of 27 isolates were found to be multi-drug resistant. Based on combination disc diffusion test and HiCrome ESBL agar method, 20 (74.07%) and 25 (92.59%) isolates were detected as ESBL producers, respectively. Among virulence genes studied, presence of wabG (25/27; 92.59%) was higher than kfuBC (5/27; 18.51%). Beta-lactamase genes viz., blaSHV, blaTEM and blaCTX-M-1 were detected in 23/27 (85.18%), 3/27 (11.11%) and 2/27 (7.40%) of isolates, respectively; while, none of the isolates was found to be positive for blaCTX-M-9 and blaOXA-1 genes. Outcome of the study provided an insight into virulence genes and ESBL producing K. pneumoniae isolated from bovine milk samples in India.
Microbiology
Ahmad Enferadi; Abdulghaffar Ownagh; Musa Tavassoli
Volume 15, Issue 2 , February 2024, , Pages 89-95
Abstract
Borrelia species are spirochetes transmitted by ticks that are important in human and animals. In most countries, there is still no molecular epidemiology of borreliosis in ruminants. This study was aimed to evaluate the existence of Borrelia spp. DNA in the blood samples of small ruminants using polymerase ...
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Borrelia species are spirochetes transmitted by ticks that are important in human and animals. In most countries, there is still no molecular epidemiology of borreliosis in ruminants. This study was aimed to evaluate the existence of Borrelia spp. DNA in the blood samples of small ruminants using polymerase chain reaction (PCR) method in West Azerbaijan Province, Iran. To detect Borrelia spp. DNA, about 1,018 ruminants (456 goats and 562 sheep) blood samples were examined from different bioclimatic regions in West Azerbaijan province, Iran. The DNA extracting and PCR were conducted. In sheep, the following prevalence rates were respectively obtained for the 16S rRNA, 5S - 23S rRNA and ospA genes: 3.55% (20/562), 2.13% (12/562) and 0.88% (5/562). And so, the prevalence rates of the genes in goats were 0.87% (4/456) for 5S - 23S rRNA gene, 1.75% (8/456) for 16S rRNA gene and 0.65% (3/456) for ospA gene. The prevalence of Borrelia spp. was significantly different in small ruminants based on the farms and localities. The sheep and goats in humid areas (north of West Azerbaijan) were infected statistically more than those in sub-humid areas (south of West Azerbaijan). It is demonstrated that host species like sheep and goats may have a key role in natural Lyme disease cycles and other borreliosis diseases in Iran.
Microbiology
Monika Punia; Sushila Maan; Kanisht Batra; Aman Kumar; Narender Singh Maan; Suresh Kumar Gahlawat
Volume 15, Issue 1 , January 2024, , Pages 13-19
Abstract
Viral and bacterial gastroenteritis and diarrhea have long been a problem in livestock with devastating effects on animal health and production causing a heavy financial burden on producers. Therefore, the bead-based multiplex detection assay was created for simultaneous detection of three livestock ...
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Viral and bacterial gastroenteritis and diarrhea have long been a problem in livestock with devastating effects on animal health and production causing a heavy financial burden on producers. Therefore, the bead-based multiplex detection assay was created for simultaneous detection of three livestock viral diarrheic agents viz. bovine rotavirus (BRV), bovine coronavirus (BCoV) and bluetongue virus (BTV). The primers and probes for triplex MAGPIX assay for simultaneous detection of three enteric viruses were designed and the assay was optimized for hybridization temperature, primer-probe and bead concentrations. The newly developed MAGPIX assay was used to determine the prevalence of these diarrhea-associated viruses by testing 200 fecal samples collected from Haryana state of India during 2018-2019. The limit of detection of the developed triplex assay was 1 × 105, 1 × 104, and 1 × 105 RNA copies for BRV, BCoV, and BTV, respectively, being lower than the reverse transcription-quantitative polymerase chain reaction (RT-qPCR). However, it was higher than the conventional RT-PCR, showing it to be more sensitive. The newly developed MAGPIX assay was a rapid, cost-effective and high throughput diagnostic tool for identification of three major entero-pathogenic diarrhea associated viruses, either alone or in tandem, with the aim to prevent and control viral diarrhea in animals.
Microbiology
Anita Dalal; Naresh Kumar Kakker; Deepika Chaudhary; Aman Kumar; Narender Singh Maan; Sushila Maan
Volume 14, Issue 12 , December 2023, , Pages 635-641
Abstract
World Organization for Animal Health has listed bluetongue (BT) under notifiable diseases. The BT is an arboviral infectious disease of domestic and wild ruminants caused by the bluetongue virus (BTV). Southern states of India had remained the point of attention for BT since first presence in 1964 in ...
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World Organization for Animal Health has listed bluetongue (BT) under notifiable diseases. The BT is an arboviral infectious disease of domestic and wild ruminants caused by the bluetongue virus (BTV). Southern states of India had remained the point of attention for BT since first presence in 1964 in Maharashtra. Recently, northern states of India have also been reported positive for BTV in small ruminants. The present study reported the dual infection of BTV serotypes, BTV-12 and -16 in sheep population from Sirsa district of Haryana in the year 2016. After detection and serotyping with Seg-2 specific real time polymerase chain reaction (PCR), the Seg-2 and Seg-6 of BTV were PCR amplified and sequenced. On phylogenetic analysis it was detected to be clustered in nucleotype G and nucleotype B specific for BTV-12 and BTV-16, respectively. This was the first report of BTV-16 from Haryana. The results signified the co-infection of two different serotypes in an animal from a single outbreak.
Microbiology
Heidar Rahimi; Amir Tukmechi; Ehsan Rashidian
Volume 14, Issue 12 , December 2023, , Pages 649-657
Abstract
The genetic diversity of Brucella strains has not been fully understood. To investigate this, the genetic characteristics of 64 isolates of Brucella melitensis from sheep and goats’ milk were studied using random fragment length polymorphism (RFLP) and multiple locus variable-number tandem repeat ...
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The genetic diversity of Brucella strains has not been fully understood. To investigate this, the genetic characteristics of 64 isolates of Brucella melitensis from sheep and goats’ milk were studied using random fragment length polymorphism (RFLP) and multiple locus variable-number tandem repeat analysis (MLVA-16) methods developed in Orsay, France (MLVA-16Orsay). The RFLP analysis revealed that all 64 isolates were of biovar one. The MLVA-typing showed that one sample was simultaneously infected with two strains of B. melitensis and the genotype of 65 isolate was analyzed. Four genotypes (47, 42, 43, and 63) were identified using MLVA-8 (panel 1), whereas six genotypes (138, 125, 116, 108, and two unknown genotypes) were identified using MLVA11 (panels 1 and 2A). From the review of MLVA-16 (panels 1, 2A, and 2B), panel 2B showed a very high discriminatory power. Two loci of Bruc04 and Bruc30 from this panel had diversity index values higher than 0.71 and the average diversity index was 0.619. So MLVA-16Orsay 34 showed the genotype indicating a low genetic homogeneity among the isolates. The findings of MLVA genotyping of the isolates suggest that strains of B. melitensis isolated from the milk of small ruminants in Iran are most closely related to the isolates from neighboring countries of the Eastern Mediterranean group. To the best of our knowledge, this is the first study to indicate the potential use of MLVA genotyping for simultaneous detection of specimen contamination using two different B. melitensis biovars.
Microbiology
Somayeh Bahrami; Mohammad Mehdi Feizabadi; Nader Mosavari; Fattah Sotoodehnejad; Mohammad Eslampanah
Volume 14, Issue 12 , December 2023, , Pages 659-664
Abstract
The new strategy for vaccine development such as the fused protein multi-epitope capable of preventing the reactivation of latent tuberculosis infection (LTBi) can be an effective strategy for controlling tuberculosis (TB) worldwide. This study was conducted to evaluate the immunity of experimentally ...
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The new strategy for vaccine development such as the fused protein multi-epitope capable of preventing the reactivation of latent tuberculosis infection (LTBi) can be an effective strategy for controlling tuberculosis (TB) worldwide. This study was conducted to evaluate the immunity of experimentally infected BALB/c mice with Mycobacterium tuberculosis after injection of DNA construct. Nineteen female BALB/c mice were divided into three groups and injected with 0.50 mL of M. tuberculosis. After 3 weeks, lung and spleen samples from the infected mice were examined. The protective effects of light chain 3-fused protein multi-epitope against TB were evaluated for post-exposure and therapeutic exposure. The lungs and spleens of the mice were aseptically removed after death for histopathology analysis. The bacterial colonies were counted, and the cells were stained after 3 weeks of incubation. No significant differences were observed between the post-exposure and therapeutic exposure groups. The pathological changes in the lung tissue of mice in these groups included an increase in the thickness of interalveolar septa, hyperemia, and intraparenchymal pulmonary hemorrhage centers (positive control), scattered hyperemic areas (negative control), and hyperemia in the interstitial tissue, scattered hyperemic areas in the lung parenchyma and lymphocytic infiltration centers (experimental group). Flow cytometry of the post-exposure and therapeutic exposure models showed insignificant changes in all three groups. It seems necessary to develop a post-exposure and therapeutic exposure vaccine strategy that focuses on LTBi to prevent the progression of the active disease. In this regard, multi-epitope vaccines should be designed to induce both cellular and humoral immunity.
Microbiology
Seyed Mahmoud Azimi; Baharak Mohammadian; Mohammad Khezri
Volume 14, Issue 12 , December 2023, , Pages 681-684
Abstract
Foot-and-mouth disease (FMD), a highly contagious viral disease of livestock, is endemic in Iran. To investigate the prevalence of antibodies against 3ABC non-structural protein (NSP) of FMD virus, a cross-sectional study was conducted on dairy cattle in eight cities of Kurdistan Province from May to ...
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Foot-and-mouth disease (FMD), a highly contagious viral disease of livestock, is endemic in Iran. To investigate the prevalence of antibodies against 3ABC non-structural protein (NSP) of FMD virus, a cross-sectional study was conducted on dairy cattle in eight cities of Kurdistan Province from May to September 2016. Serum samples (n = 283), were collected from cattle vaccinated with the recommended dose of a commercial vaccine and tested by a Competition enzyme-linked immunosorbent assay. Results showed the overall seroprevalence of antibodies against NSP of FMD virus in the vaccinated cattle was 22.30% (95.00% CI: 17.40 - 27.20%). The seroprevalence of antibodies was affected by geographical regions, with the highest seroprevalence related to the samples of vaccinated cattle in the cities of Marivan 95.00% (95.00% CI: 92.50 - 97.50%) and Saqqez 38.50% (95.00% CI: 32.80 - 44.20%). In terms of age, the highest seroprevalence of antibodies to FMD virus 26.70% (95.00% CI: 21.60-31.80%) belonged to ≤ 24-month-old cattle. These findings suggest that the presence of NSP antibodies in vaccinated cattle indicates the risk of infection with FMD virus serotypes circulating in the west of the province, so further studies with a larger sample size are recommended.
Microbiology
Yongxiang Liu; Lide Qin; Xiaoliang Hu; Yanmei Jiang
Volume 14, Issue 11 , November 2023, , Pages 583-588
Abstract
Feline calicivirus (FCV) is a highly contagious pathogen seriously affecting the upper respiratory tract and producing oral diseases in the feline. Despite widespread vaccination, the prevalence of FCV remains high. In this study, the FCV qingdao (qd)/2019/china was isolated from a domestic feline oropharyngeal ...
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Feline calicivirus (FCV) is a highly contagious pathogen seriously affecting the upper respiratory tract and producing oral diseases in the feline. Despite widespread vaccination, the prevalence of FCV remains high. In this study, the FCV qingdao (qd)/2019/china was isolated from a domestic feline oropharyngeal swab collected from Qingdao, China. The virus was purified using the plaque assay and identified using the Polymerase chain reaction and indirect immunofluorescence assay methods, the capsid amino acid, VP1 of qd/2019/china, showed sequence identity with the other isolates ranging between 83.90% (ym3/2001/jp) and 91.10% (CH-JL4). The sequence of the capsid amino acid revealed qd/2019/china to be closely related to CH-JL4 and clustered with CH-JL4 in the phylogenetic tree. The phylo-genetic analysis indicated that the complete genomes (GenBank® accession No. MZ322896) of qd/2019/china and CH-JL4 were also classified into the same cluster. The recombination analysis with Simplot indicated that the qd/2019/china originated from the recombination of CH-JL4 and HRB-SS, and the region 3,821 - 5,301 nt originated from HRB-SS. Further, the region 3,821 - 5,301 nt were found to belong to the protease-polymerase (PP) of HRB-SS. Here, we isolated a new recombinant virus, FCV qd/2019/china. Therefore, these results would be beneficial for better understanding of the evolution and epidemiology of FCV.
Microbiology
Mehrnoosh Gadir; Seyed Mahmoud Azimi; Naser Harzandi; Behzad Hemati; Neda Eskandarzade
Volume 14, Issue 11 , November 2023, , Pages 615-623
Abstract
Despite widespread vaccination against foot-and-mouth disease, many outbreaks still occur in endemic areas. We attempted to determine the genetic and antigenic properties of the O/PanAsia-2/QOM-15 foot-and-mouth disease virus new vaccine strain. Thus, whole-genome sequencing was used to identify vulnerable ...
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Despite widespread vaccination against foot-and-mouth disease, many outbreaks still occur in endemic areas. We attempted to determine the genetic and antigenic properties of the O/PanAsia-2/QOM-15 foot-and-mouth disease virus new vaccine strain. Thus, whole-genome sequencing was used to identify vulnerable pinpoint sites across the genome. The VP1 sequence (1D gene) of the O/PanAsia-2/QOM-15 viral genome was then compared to the VP1 sequences of two previously used vaccine strains, O/PanAsia (JQ321837) and O/PanAsia-2 (JN676146). The antigenic relationship of these three viruses was calculated by the two dimensional-virus neutralization test. At the nucleotide level, 47 single variants were identified, of which 19.00% were in the 5' untranslated region (UTR), 79.00% in the polyprotein region, and 2.00% in the 3' UTR region. Approximately half of the single nucleotide polymorphisms that have occurred in 1D gene resulted in amino acid (AA) substitutions in the VP1 structure. The single nucleotide polymorphisms also caused AA substitutions in other structural proteins, including VP2 and VP3, and some non-structural proteins (Lpro, 2C, and 3A). The O/PanAsia-2/QOM-15 shared higher sequence similarity with O/PanAsia-2 (91.00%) compared to O/PanAsia (87.30%). Evaluating r-value showed that the antigenic relationship of O/PanAsia-2/QOM-15 with O/PanAsia-2 (29.00%) was greater than that of the O/PanAsia (24.00%); however, all three viruses were immunologically distinct. After 10 years, the alteration of virus antigenicity and the lack of detectable adaptive pressure on VP1 sequence suggest that studying genetic dynamics beyond the VP1 region is necessary to evaluate FMDV pathogenicity and vaccine failure.
Microbiology
Mohammad Tabatabaei; Fateme Abdolahi
Volume 14, Issue 9 , September 2023, , Pages 481-487
Abstract
Pasteurella multocida exists as a commensal in the upper respiratory tracts of livestock, and poultry, and causes a wide variety of diseases in humans and animals. This study aimed to investigate the incidence of P. multocida by bacteriological and molecular characterization in sheep and goats and screening ...
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Pasteurella multocida exists as a commensal in the upper respiratory tracts of livestock, and poultry, and causes a wide variety of diseases in humans and animals. This study aimed to investigate the incidence of P. multocida by bacteriological and molecular characterization in sheep and goats and screening the existence of capsule-specific genes and their antibiotic resistance pattern. Totally, 1650 nasopharyngeal swabs were collected from apparently healthy sheep and goats and 460 lung tissues were collected from slaughtered animals in Fars province, Iran. All samples were cultured and suspected colonies were examined by biochemical tests, antimicrobial assay and polymerase chain reaction (PCR). Among 165 P. multocida (104 sheep and 61 goats) isolates, the capA, capD, and capB genes were amplified in 98, 48, and 12 isolates, respectively. The occurrence of four virulence-associated genes of P. multocida isolates were determined by PCR. Most isolates harbored the toxA (79.40%) and hgbB genes (70.90%) and 59.40% of isolates had the pfhA gene. Almost half of the isolates (46.10%) contained the tbpA gene. According to the current study, P. multocida capsular type A had the most frequency followed by type D. In addition, the high frequency of tbpA, pfhA, toxA, and hgbB genes revealed that these genes are possibly important in the pathogenesis of P. multocida. Oxytetracycline, enrofloxacin, florfenicol, and tilmicosin were the most effective drugs.
Microbiology
Razieh Sadati; Nima Shaykh-Baygloo; Rasoul Shokri
Volume 14, Issue 9 , September 2023, , Pages 515-523
Abstract
Isolation of new microbial species from extreme environments is one of the most efficient approaches for the development of novel bioactive metabolites. The aim of the present study was to explore the pharmaceutical bacterial resources from the water and sediments of hypersaline Lake Urmia. Using different ...
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Isolation of new microbial species from extreme environments is one of the most efficient approaches for the development of novel bioactive metabolites. The aim of the present study was to explore the pharmaceutical bacterial resources from the water and sediments of hypersaline Lake Urmia. Using different culture conditions and media led to the isolation of 20 bacterial strains. Halophilic bacteria were screened for the production of antibacterial agent against multi-drug resistant strains of Escherichia coli through agar well diffusion assay. Halophilic bacteria DNA extraction was done by boiling method. The results showed that two Halomonas strains, LUH16 and LUH20 identified by analysis of 16S rRNA gene sequences were the potent producers of antimicrobial metabolites against various strains of E. coli. Furthermore, gas chromatography-mass spectrometry (GC-MS) analysis revealed the presence of eight secondary metabolites with the relevant antimicrobial properties. Our findings led us to focus on Halomonas strains as potent producers of antimicrobial compound that might be an alternative against antibiotic-resistant pathogens such as pathogenic Escherichia coli.
Microbiology
Seyed Sajjad Babaeimarzangou; Manoochehr Allymehr; Aidin Molouki; Alireza Talebi; Mohammad Hossein Fallah Mehrabadi
Volume 14, Issue 8 , August 2023, , Pages 447-456
Abstract
Newcastle disease virus (NDV) is considered one of the most devastating avian viral patho-gens affecting the avian population, and it causes a significant economic burden on the poultry industry worldwide. The study aimed to gain deeper understanding of the molecular and phylogenetic analyses of the ...
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Newcastle disease virus (NDV) is considered one of the most devastating avian viral patho-gens affecting the avian population, and it causes a significant economic burden on the poultry industry worldwide. The study aimed to gain deeper understanding of the molecular and phylogenetic analyses of the complete hemagglutinin-neuraminidase (HN) coding region among NDV isolates. The samples were obtained from different parts of Iran from July 2017 to February 2020, were used for phylogenic analysis in this study. The results confirmed the predominance of sub-genotype VII.1.1, previously known as sub-genotype VIIL, which is circulating in commercial broiler farms of Iran. Identification of (a) an additional N-glycosylation site (NIS) at position 144; (b) mutations S315P and I369V which are related to increasing the viral thermostability; (C) cysteine residues at positions 123; (d) amino acid substitutions in the HN antigenic sites, especially the mutations I514V and E347Q, as well as the other mutant within HN binding sites of the VII.1.1 sub-genotype, suggests the idea that this new sub-genotype of NDV may possess a high level of pathogenicity and virulence compared to other NDV sub-genotypes. In conclusion, the results indicate the presence of an additional NIS at position 144, which may alter the virulence of the isolates. Furthermore, the presence of the thermostable mutations (S315P and I369V) and the other amino acid substitutions among the VII.1.1 sub-genotype isolates may have an impact on the vaccine immunity against this new NDV sub-genotype.
Microbiology
Dharitree Sonowal; Sandeep Ghatak; Acheenta Gohain Barua; Srinivas Kandhan; Razibuddin Ahmed Hazarika; Arnab Sen; Samir Das; Sarat Sonowal; Rajeev Kumar Sharma; Shantanu Tamuly; Chimanjita Phukan; Ajanta Sharma; Poznur Hussain
Volume 14, Issue 7 , July 2023, , Pages 351-358
Abstract
Staphylococcus aureus are Gram positive bacteria known to acquire antibiotic resistance rapidly and pose a major challenge to clinicians worldwide. Infections by methicillin resistant Staphylococcus aureus (MRSA) are usually associated with increased mortality and prolonging of treatment. Samples (n ...
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Staphylococcus aureus are Gram positive bacteria known to acquire antibiotic resistance rapidly and pose a major challenge to clinicians worldwide. Infections by methicillin resistant Staphylococcus aureus (MRSA) are usually associated with increased mortality and prolonging of treatment. Samples (n = 706) from diverse sources (livestock, pets, animal handlers, human hospital) were collected and screened for the presence of MRSA by phenotypic and genotypic methods. The incidence of Staphylococcus aureus was greater in goats (42.00%; 28.20 - 56.80%, confidence interval [CI] 95.00%) followed by cattle (13.50%; 9.20 - 18.80%, CI 95.00%), humans (12.90%; 9.30 - 17.40%, CI 95.00%) and dogs (12.90%; 8.10 - 19.20%, CI 95.00%). Significantly higher incidence of MRSA was observed in dogs (65.00%; 40.80 - 84.60%, CI 95.00%), compared to other hosts namely cattle (48.00%; 26.50 - 64.30%, CI 95.00%), humans (35.00%; 20.20 - 52.50%, CI 95.00%) and goats (10.00%; 1.20 - 30.40%, CI 95.00%). All the S. aureus isolates were further screened for thermostable nuclease (nuc gene) by polymerase chain reaction (PCR). The incidence of nuc gene in cattle, dog, goat and human were found to be 3.30% (1.30 - 6.60%, CI 95.00%), 5.20% (2.30 - 9.90%, CI 95.00%), 28.00% (16.20 - 42.50%, CI 95.00%) and 9.10% (6.00 - 13.00%, CI 95.00%), respectively. Comparative evaluation of two PCR primers (mecA-162 and mecA-310) indicated the former one as more rational choice for detection of MRSA. Overall, the results of our study indicated possible risk of zoonotic transmission of MRSA from canines.
Microbiology
Maryam Dadar; Saeed Alamian; Ali Mohammad Behrozikhah; Freshteh Yazdani; Armin Kalantari; Afshar Etemadi; Adrian M. Whatmore
Volume 10, Issue 4 , December 2019, , Pages 315-321
Abstract
Brucellosis is a costly contagious disease of human, domestic and wild animals. It is a serious health problem in Iran causing significant economic losses therefore, control approaches to prevent its spread are of great importance. In Iran, the species and biovars of virulent Brucella species are still ...
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Brucellosis is a costly contagious disease of human, domestic and wild animals. It is a serious health problem in Iran causing significant economic losses therefore, control approaches to prevent its spread are of great importance. In Iran, the species and biovars of virulent Brucella species are still under-reported due to the inadequate diagnostic protocols and insufficient laboratory facilities. The objective of this study was to characterize Brucella isolates obtained from passive animal and human surveillance in Iran from 2011 to 2018 in order to understand the current epidemiological situation of the disease. A total of 419 samples (milk, blood, cerebrospinal fluid, abomasum content and aborted fetus tissues) were collected from 65 cases/case series (human and animals) and examined bacteriologically. The initially identified Brucella isolates were further characterized using phenotypic and molecular approaches. All recovered isolates were either B. abortus or B. melitensis. The infection in sheep appeared to be exclusively associated with B. melitensis, but both B. abortus and B. melitensis were common in bovine samples. Samples from one sheep and one goat were confirmed to be infected by the B. melitensis vaccine strain Rev1. In spite of B. abortus burden in animals (14 cases in cattle and camel), brucellosis in human was predominantly associated with B. melitensis (15 cases). The results confirmed that B. melitensis biovar 1 and B. abortus biovar 3 remain the most prevalent biovars in Iran. This report builds a picture of the significance of different Brucella species in different hosts in Iran and provides applicable information for the healthcare professionals about the public health risks of brucellosis and relevant preventive strategies.
Microbiology
Meryem Cansu Yesiltas; Ilhan Altinok; Rafet Cagri Ozturk
Volume 10, Issue 2 , June 2019, , Pages 101-107
Abstract
Lactococcosis disease incident caused by Lactococcus garvieae has been increased with increasing aquaculture productions and outbreaks of the disease have become a threat on farmed species. To prevent lactococcosis, inactivated vaccine has been used, however, it only provides protection when given by ...
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Lactococcosis disease incident caused by Lactococcus garvieae has been increased with increasing aquaculture productions and outbreaks of the disease have become a threat on farmed species. To prevent lactococcosis, inactivated vaccine has been used, however, it only provides protection when given by injection. Other than inactivated vaccine, various vaccines such as subunit vaccines can be developed. In the present study, total protein profile of 43 strains of L. garvieae isolated from fish, milk and cheese by SDS-PAGE and virulence associated immunogenic proteins of L. garvieae strains using western blot with hyper-immune rabbit sera were determined. After analyzing whole-cell lysate protein of L. garvieae strains with SDS-PAGE, protein bands were ranged between 8.00 and 140.00 kDA. Among strains, variable protein bands were ranged between 17.00 and 48.00 kDa with some variability in the staining intensity of the protein bands and formed in 6 clusters. The immunogenic protein bands were ranged between 25.00 - 75.00 kDa. Only a variable and highly immunogenic protein band was observed between 40.00 and 45.00 kDa. Most of the strain including Lgper had 44.00 kDa immunogenic protein while nonvirulent ATCC strain had 42.50 kDA immunogenic protein. Predominant immuno-reactive proteins encoded by genes can be used as a subunit vaccine.
Poultry
Guang Zhao; Tingting Huang; Di Wu; Liwu Zhang; Zeli Luo; Jia Liu; Xiaowei Yang
Volume 10, Issue 2 , June 2019, , Pages 169-172
Abstract
Duck beak atrophy and dwarfism syndrome (BADS) is a newly emerged disease in China since 2015. In October 2017, an unidentified disease occurred in Cherry Valley ducks, Chongqing municipality, the southwest of China. The affected birds showed short beak and growth retardation clinical signs. The disease ...
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Duck beak atrophy and dwarfism syndrome (BADS) is a newly emerged disease in China since 2015. In October 2017, an unidentified disease occurred in Cherry Valley ducks, Chongqing municipality, the southwest of China. The affected birds showed short beak and growth retardation clinical signs. The disease caused approximately 20.00% morbidity and serious weight loss due to retarded growth. In order to identify the causative agent of BADS, liver, spleen, lung and heart samples were collected for virus isolation, hemagglutination test, PCR identification, and partial gene sequencing. The isolated virus was tentatively named SC16. Hemagglutination test indicated that the virus was negative to chicken red blood cells. Based on the PCR and sequencing results, the causative agent of BADS was a novel duck-origin goose parvovirus (DGPV) while no another co-infection pathogen was found in this case. Further analysis could provide insights into the control strategies of DGPV in ducks.
Microbiology
Siavash Maktabi; Masoud Ghorbanpoor; Masomeh Hossaini; Amirabbas Motavalibashi
Volume 10, Issue 1 , March 2019, , Pages 37-42
Abstract
Campylobacter jejuni and C. coli are the main causes of gastrointestinal diseases in humans even in industrialized countries affecting public health. The aim of the current study was to evaluate the occurrence and antibiotic resistance of C. jejuni and C. coli in chicken meat, beef, mutton and water ...
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Campylobacter jejuni and C. coli are the main causes of gastrointestinal diseases in humans even in industrialized countries affecting public health. The aim of the current study was to evaluate the occurrence and antibiotic resistance of C. jejuni and C. coli in chicken meat, beef, mutton and water buffalo meat slaughtered in Ahvaz city, Iran. A total of 380 samples including chicken meat from industrial abattoirs (n = 150), chicken meat from traditional abattoirs (n = 50), fresh packed chicken meat from local markets (n = 30) and beef, mutton and water buffalo meat from industrial abattoirs (50 samples for each meat) in Ahvaz,were collected and tested for the presence of Campylobacter spp. The procedure was one-step enrichment in Preston enrichment broth followed by plating on supplemented blood agar for 24 hr under microaerophilic conditions at 42 ˚C. Suspected colonies were tested by polymerase chain reaction assay and susceptibility of the confirmed isolates to various antibiotics was investigated by the Kirby-Bauer disk diffusion method. Overall, 32 samples (8.40%) were contaminated with Campylobacter spp. Mutton was the most contaminated meat (24%), while fresh packed chicken meat were not contaminated. Among the 32 isolates, 40.60%, 34.40%, 21.90%, and 15.60% were resistant to tetracycline, ciprofloxacin, ampicillin, and streptomycin, respectively. Moreover, a high number of multi-antibiotic resistant Campylobacter spp. was determined. Since foods of animal origin are the most sources of Campylobacter infection, the presence of resistant strains to antibiotics is a potential risk to public health.
Microbiology
Soheila Najafi; Morad Rahimi; Zahra Nikousefat
Volume 10, Issue 1 , March 2019, , Pages 43-49
Abstract
Pathogenic Escherichia coli strains cause a wide range of extra intestinal infections including urinary tract infection in humans and colibacillosis in poultry. They are classified into uropathogenic E. coli (UPEC) and avian pathogenic E. coli (APEC) with genetic similarities and variations. Their ...
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Pathogenic Escherichia coli strains cause a wide range of extra intestinal infections including urinary tract infection in humans and colibacillosis in poultry. They are classified into uropathogenic E. coli (UPEC) and avian pathogenic E. coli (APEC) with genetic similarities and variations. Their pathogenicity is related to the virulence-encoding genes like sfa, papG II, ompT, iutA, and iss with zoonotic potentials. One hundred isolated E. coli from patients with urinary tract infection and 100 E. coli from chickens with colibacillosis were evaluated for the presence of the most common virulence-encoding genes including sfa, papG II, ompT, iutA, and iss by multiplex polymerase chain reaction. While the frequency of sfa, papG II, ompT, iutA and iss encoding genes in APEC isolates were respectively 0.00%, 67.00%, 63.00%, 89.00% and 89.00%, the frequency of these encoding genes in UPEC isolates were 18.00%, 40.00%, 40.00%, 74.00% and 48.00%, respectively. Except for sfa, the frequencies of other encoding genes in APEC were more than those in UPEC isolates. The iutA as the most common UPEC encoding gene and iss as the most common APEC encoding gene were the most prevalent virulence factors in the examined E. coli isolates. Finding out the distribution of virulence-associated genes could be helpful to identify similarities and differences between APEC and UPEC isolates in order to provide more substantial evidence of their common virulence traits and potential zoonotic threats.
Microbiology
Jamshid Razmyar; Mahdis Ghavidel; Hamideh Salari Sedigh
Volume 10, Issue 1 , March 2019, , Pages 67-72
Abstract
Genus Brachyspira,as Gram negative anaerobic bacteria, colonize in dogs intestine. The aim of the current study was to determine the prevalence of Brachyspira spp. for the first time in Iran and rapid identification of Brachyspira spp. in dogs by a new designment of a species-specific primer set for ...
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Genus Brachyspira,as Gram negative anaerobic bacteria, colonize in dogs intestine. The aim of the current study was to determine the prevalence of Brachyspira spp. for the first time in Iran and rapid identification of Brachyspira spp. in dogs by a new designment of a species-specific primer set for B. canis. One hundred fifty-one fecal samples were obtained from dogs by rectal swab. Twenty dogs suffered from diarrhea and 131 of them were healthy. In 9.27% (14/151) of samples, spirochaetes were detected on primary cultures by weak hemolysis and positive Gram staining and then Brachyspira genus was confirmed by NADH oxidase (nox) gene via polymerase chain reaction. Among 14 isolates, twelve isolates were B. canis, one isolate was B. intermedia and another one was non-typeable. From 12 B. canis, only eight isolates were detected by designed specific primers. Ten Brachyspira spp. were isolated from dogs ≤ 1 year old (10/67, 14.92%) and 4 isolates were from > 1 year old dogs (4/84, 4.76%). The isolation rates from healthy and diarrheic dogs were (12/131, 9.16%) and (2/20, 10.00%), respectively. A statistically significant association was observed between the presence of Brachyspira spp. and the age under one year. Based on our findings, the nox gene in B. canis might have more sequence variability compared to other Brachyspira spp.
Microbiology
Zahra Yadegari; Gholamreza Nikbakht Brujeni; Reyhaneh Ghorbanpour; Farhad Moosakhani; Samad Lotfollahzadeh
Volume 10, Issue 1 , March 2019, , Pages 73-78
Abstract
Enterotoxigenic Escherichia coli (ETEC) produces different virulence factors allowing the bacterium to colonize and develop watery diarrhea. Proteomics studies have also introduced new protein belonging to the secretion pathways, antigen 43 (Ag43), which plays important role in E. coli pathogenesis. ...
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Enterotoxigenic Escherichia coli (ETEC) produces different virulence factors allowing the bacterium to colonize and develop watery diarrhea. Proteomics studies have also introduced new protein belonging to the secretion pathways, antigen 43 (Ag43), which plays important role in E. coli pathogenesis. The objective of this study was to investigate O-types and virulence factors of E. coli isolates from neonatal calves diarrhea. Total of 120 isolates from diarrheic calves were genotyped for their O groups and the presence of virulence genes K99, F41 and STa as well as Ag43. The predominant O-type was O101 (51.00%) and the prevalence of K99, F41 and STa was 7 (5.80%). The Ag43 was detected in all samples with three different allelic patterns. Our results indicated that K99 positive isolates certainly have one of each 2200 bp or 1800 bp or both copies of Ag43 passenger domain, while negative K99 isolates lack the Ag43. The results reported here provide informative data regarding the prevalence of E. coli O-types and their virulence factors in enteric colibacillosis. The Ag43 that was more found in K99 positive isolates might be associated with diarrhea-causing E. coli strains in neonatal calves.
Microbiology
Javad Aliakbarlu; Sindokht Ghiasi; Behnaz Bazargani-Gilani
Volume 9, Issue 4 , December 2018, , Pages 361-365
Abstract
Barberry fruit is consumed in different forms including dried fruit, juice, jam and marmalade in Iran. This fruit is also used as a food additive (flavoring and colorant) in soup and rice dishes. In present study, antioxidant activities of acetone, ethanol and water (infusion and decoction) extracts ...
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Barberry fruit is consumed in different forms including dried fruit, juice, jam and marmalade in Iran. This fruit is also used as a food additive (flavoring and colorant) in soup and rice dishes. In present study, antioxidant activities of acetone, ethanol and water (infusion and decoction) extracts of barberry (Berberis vulgaris) fruit were investigated using 2,2-azinobis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and reducing power methods. Total phenolic contents of the extracts were also estimated using Folin-Ciocalteu assay. In ABTS assay, acetone and ethanol extracts showed the highest radical scavenging activity, while in DPPH and reducing power methods, acetone extract and decoction exhibited the strongest antioxidant activity. Meanwhile, the antioxidant potential of water extracts increased with increasing heating time (antioxidant activity of decoction was higher than that of infusion). The highest total phenolic content was found in the acetone extract (92.75 mg GAE per g). It was concluded that the acetone extract and decoction of barberry fruit can be used as an effective natural antioxidant in food industry.