Synthesis and Evaluation of the Immunomagnetic Beads for Separation of the Salmonella typhimurium from Milk.

Salmonella causes zoonotic diseases in humans and many animal species. The bacteria could be spread through fecal-oral transmission and consumption of raw contaminated animal products. Despite the activities which are carried out for the prevention of salmonellosis, it causes economic losses. This study aimed to prepare immunomagnetic beads to separate the Salmonella bacteria from experimentally polluted milk samples. The antibodies were purified from the rabbit's hyperimmune sera and coupled to the Fe nanoparticles using diethylenetriaminepentaacetic acid (DTPA) as a linker. The synthesized particles were analyzed using electron microscopy. The limit of bacterial detection by using the immunomagnetic beads coupled with bacterial culture were tested in experimentally contaminated cow milk with Salmonella. The separated bacteria were identified by using bacterial culture and biochemical tests. Using immunomagnetic beads (IMB), the Salmonella bacteria were removed from milk samples, concentrated in sterilized PBS, and cultured in nutrient agar media. The conventional culture method detected the bacteria in samples polluted with at least 3×104 CFU/mL bacteria; however, isolated bacteria were separated from milk samples using IMB and defined on bacterial culture media. The 3 CFU/mL of S. Typhimuriumm were detected in experimentally polluted milk samples using the current immunomagnetic-culture method. The results suggested using the IMB-bacterial culture instead of the conventional culture method.

Effects of Selected Adjuvants on Immunogenicity and Protectivity of Pasteurella multocida Bacterin Vaccine in Chickens.

Avian pasteurellosis (fowl cholera) is an important disease affecting domestic and wild birds all over the world. Although the capsular type A of Pasteurella multocida is mostly involved, other capsular types are occasionally incriminated. The present study aimed at investigating the effect of some adjuvants on immunogenicity and protectivity of P. multocida bacterin in chickens, compared to an Iranian commercial vaccine. Eight-week-old chicken pullets were double vaccinated with an interval of three weeks. Vaccine immunogenicity testing was conducted using an in-house indirect enzyme-linked immunosorbent assay and assessing serum antibody titers at 7, 14, and 21 days post-primary and 14 days post-secondary immunization. The possible adverse effects were recorded by a poultry-disease expert. For evaluating the vaccine protection rate, chickens were subjected to 2×Lethal Dose 50%of a virulent P. multocida strain two weeks post-secondary immunization. The rate of live and normal animals was regarded as protection rate 7days after the exposure. The findings showed that oil adjuvants Montanide ISA 70-and Montanide ISA 71-containingvaccines (with or without saponin) caused a powerful immune reaction than the aluminum adjuvanted vaccine and commercial vaccine (P<0.05). Significant protection against challenge was merely induced by the oil adjuvanted vaccines (P<0.05). The majority of the studied chickens showed inflammation at the injection site (yellow) throughout the trial. Vaccines made by Montanide ISA 70 and Montanide ISA 71 are novel and effective inactivated vaccines that are able to cause significant protection to fowl cholera disease.

Restriction Fragment Length Polymorphism Typing of Staphylococcus aureus Strains Isolated from Bovine Mastitis and Dairy Products in Ahvaz, Iran, Using of Digested Coagulase Gene.

Staphylococcus aureus is a major pathogen in the transmission of diseases from animals to humans and vice-versa.Various infections, such as mastitis in cattle, sheep and goats, as well as gastroenteritis due to food poisoning in humans are the most frequent problems caused by S. aureus. The bacteria also lead to severe economic losses in dairy industry. A major virulence factor for the organism is encoded by the coagulase (coa) gene. This study aimed to assess the polymorphisms of the coa gene in S. aureus strains isolated from bovine mastitis and dairy product samples in Ahvaz, Iran. The results showed that out of 91 S. aureus, 80 (87.91%) isolates were positive for coa gene(s). In total, nine different polymerase chain reaction (PCR) products were obtained for coa-positive isolates. A single band was detected in coa PCR with a size ranges from 370 to 830 bp in most isolates (n=77, 96.25%). For three isolates (3.75%), two amplification products were obtained. A PCR product of an estimated size of 590 bp was most frequent, as obtained for 48 (60.00%) isolates. Whereas, 370 and 830 bp PCR products were the least presented, for two (2.50%) and one (1.25%) isolate, respectively. Subsequently, for restriction fragment length polymorphism (RFLP), typing of coa gene and AluI restriction enzyme were used for the digestion of the products. AluI for most of PCR products generated a unique pattern; however, four PCR products (the sizes ranged 750, 670, 590, and 510 bp) generated three or more patterns. Based on AluI RFLP of coa gene, the isolates were classified into 23 groups. Two groups of isolates were dominant, making 45% of the total. According to the findings, one or two types of coa RFLP were dominant among samples that were infected with more S. aureus isolates belonging to different coa RFLP types.

Detection of Mycoplasma gallisepticum and Mycoplasma synoviae among Commercial Poultry in Khouzestan Province, Iran.

Mycoplasmas are important avian pathogens, which can cause both respiratory disease and synovitis in poultry that result in considerable economic losses to the poultry industry all over the world. The aim of this study was to determine the prevalence of Mycoplasma gallisepticum and Mycoplasma synoviae infections among commercial poultry flocks in Khouzestan province, Iran, using the polymerase chain reaction (PCR) technique. Totally, 290 tracheal swab samples were collected from 19 broiler flocks and 4 layer-breeder flocks, with or without respiratory signs, in different areas of Khouzestan province within six months. The PCR tests were applied for the specific amplification of 16S rRNA (185 bp) and vlhA (392 bp) genes. Out of 100 swab samples obtained from the layer-breeder flocks, 1 and 72 specimens were positive for M. gallisepticum and M. synoviae, respectively. In this regard, out of the 4 layer-breeder flocks, 1 (25%) and 4 (100%) flocks were positive for M. gallisepticum and M. synoviae, respectively. However, none of the studied broiler flocks were M. gallisepticum- or M. synoviae-positive. According to the results, the PCR technique could be concluded as a rapid method for the accurate identification of M. gallisepticum and M. synoviae infections in commercial poultry flocks. The results were indicative of the low prevalence of M. gallisepticum in the studied flocks in Khouzestan province. On the other hand, M. synoviae was widely distributed among layer-breeder flocks in this province.

Occurrence of purulent mandibular and maxillary osteomyelitis associated with Pseudomonas aeruginosa in a sheep flock in south-west of Iran.

This report describes an outbreak of purulent mandibular and/or maxillary osteomyelitis caused by Pseudomonas aeruginosa in a sheep flock located in the Khuzestan province, Iran. Jaw bones of almost 100 out of 500 mature sheep in a flock became severely deformed with a variably sized firm swelling, without any signs of inflammation in the surrounding soft tissues. The affected animals showed anorexia, depression, swelling of the mandibular and/or maxillary area, loss of cheek teeth and poor body condition. These animals were gradually culled in a period of 3 months. Postmortem examination showed a hard swelling of jaw bones with dirty greenish pus that filled alveolar molar teeth cavities. Histopathologic findings revealed necrotic areas surrounded by mixed population of inflammatory cells with exuberant fibrosis around some area of the lesions and irregular trabeculae of woven bone. In bacteriology, pure culture of P. aeruginosa was isolated from all of 7 sampled sheep. Based on clinical examination, radiography, histopathological features and bacteriology, the lesions were diagnosed as chronic suppurative osteomyelitis caused by P. aeruginosa. According to bacteriological results, the likely source of bacterial infection in this study was drinking water.

An Investigation into Enterobacteriaceae Responsible for Early Mortality in Japanese Quail Chicks and Their Antibiotic Susceptibility Patterns.

Quail is an alternative source of protein for humans. These birds can be affected by common bacterial infections. Bacterial contamination of egg is the most common cause of mortality in Japanese quail chicks. In order to study the role of some members of Enterobacteriaceae responsible for early mortality in Japanese quail chicks, 100 dead or moribund quail chicks were obtained from 10 different farms in Ahvaz, Iran. Samples were taken from the liver and yolk sac of the birds and bacterial isolation from samples was conducted by streaking them on MacConkey, Brilliant Green, Salmonella-Shigella and Xylose Lysine Deoxycholate agar plates. The plates were incubated at 37 &deg;C for 24-48 hours, and by standard biochemical tests bacterial isolates were identified. Final confirmation of Salmonella serotypes was performed by Razi Institute. All the isolates were examined for susceptibility to 12 different antibiotics (Padtan-Teb Co., Tehran, Iran) by the disk diffusion (Kirby Bauer) method. The results showed that 78% of the quail chicks were infected. The isolated bacteria were Escherichia coli (44%), Klebsiella pneumonia (8%), Salmonella serovar ruzizi (5%), Salmonella serovar typhimurium (3%), Enterobacter cloacae (4%), Enterobacter aerogenes (4%), Proteus vulgaris (5%) and Proteus mirabilis (5%). One hundred percent susceptibility was observed to gentamycin, soltrim, tetracycline, fosfomycin, florfenicol, cephalexin and ceftriaxone. E. coli isolates were susceptible to soltrim and ceftriaxone, Salmonella isolates were susceptible to fosfomycin, Enterobacter isolates were susceptible to ceftriaxone and Proteus and Klebsiella isolates showed susceptibility to ceftriaxone. It is concluded that the members of Enterobacteriaceae family, specifically the genera Escherichia and Salmonella, are the major causes of early mortality in newly-hatched Japanese quail chicks.

Testicular lesions and epididymal sperm parameters in the Iranian river buffalo (Bubalus bubalis).

The aim of this study was to evaluate the testicular lesions and their effects on the epididymal sperm parameters in the Iranian river buffalo (Bubalus bubalis). Total numbers of 117 scrota from the pubertal buffalo were provided from the local slaughterhouse. The samples were evaluated for morphological parameters and any macro- or microscopic lesions. The sterile swabs from the testis parenchyma were subjected to microbiology culture. The epididymal spermatozoon was analysed for concentration, progressive motility and abnormalities. The results showed 34.2% fibrotic adhesions between parietal and visceral layers of tunica vaginalis that was significantly different among seasons (P < 0.05). The cases of unilateral cryptorchidism and bilateral Sertoli cell tumour were detected, with no spermatozoa in the respected epididymides. Microscopic examination showed 13.25% (31/234) lesions including general (51.61%; 16/31) and multifocal (29.03%; 9/31) degenerations as well as interstitial orchitis (9.68%; 3/31) and the Sertoli cell tumour (6.45%; 2/31). No relationship between the lesions and the bacterial isolation (n = 6) was detected. The sperm parameters and morphological parameters of the testis were under influence of microscopic lesions (P < 0.05). In conclusion, the testicular macro- and microscopic lesions may have a noticeable contribution in the Iranian buffalo fertility.

Evaluation of immunopathologic effects of aqueous extract of Echinacea purpurea in mice after experimental challenge with Pasteurella multocida serotype A.

In order to assess the immunopathological effects of aqueous Echinacea purpurea extract (EPE) on mice experimentally challenged with Pasteurella multocida serotype A, forty female BALB/c mice were randomly divided into four groups. The groups included a control group (received sterile distilled water 2 times/week for 2 weeks, intraperitoneally and then 100 µl sterile saline intranasally), a PMA group (received sterile distilled water as the control group and after 2 weeks, 5.6 × 10(3) CFU/ml of P. multocida serotype A, intranasally), an EPE+PMA group (received E. purpurea extract intraperitoneally 2 times/week for 2 weeks and then challenged as the PMA group) and an EPE group (received E. purpurea extract as EPE+PMA group and then 100 µl sterile saline intranasally). After 24 and 48 h post challenge, half of the animals in each group were sacrificed and analyzed for bacterial counts in their lungs and livers, TNFα serum levels and histapathological changes. The results showed significant differences in lung bacterial counts between PMA and EPE+PMA groups. TNFα serum level was significantly higher in the PMA group. Histopathological examination revealed infiltration of neutrophils in alveolar septa and hyperemia in the PMA group. In addition, the criteria of bronchopneumonia were partially recovered in the EPE+PMA compared to the PMA group. According to the results, it seems that E. purpurea extract has an immunomodulatory effect and can be used to prevent or control of pneumonia caused by Pasteurella.