BACKGROUND: Microencapsulation of probiotic bacteria is an area that has rapidly expanded over the recent years. It is one of the new methods of improving probiotics stability, through which the biological agents are protected with an enclosed coating to release the active agents within the coating at a controlled rate in time and under special conditions.OBJECTIVES: The present study the aims to evaluate the effect of Lactobacillus plantarum microencapsulation with chitosan/alginate microparticles on the oxidative response of Nile tilapia fish (Oreochromis niloticus).METHODS: Herein, 240 pieces of Nile tilapia, with an average weight of 15.56±0.02 g, were randomly divided into four groups as follows: group1 or control with a diet without probiotics, groups 2, 3, and 4 respectively with a diet containing 108 Log CFU/g of unencapsulated Lactobacillus plantarum, a diet containing microencapsulated Lactobacillus plantarum, and with feed containing alginate with chitosan without the presence of bacteria. They were fed for 60 days at the rate of 2 % of body weight twice a day.RESULTS: The activity level of oxidative enzymes, biochemical factors, and liver enzymes of the Nile tilapia fish were investigated. The findings revealed that oxidative enzymes, biochemical factors, and liver enzymes were positively affected in the groups with microencapsulated Lactobacillus plantarum (P<0.05). The three enzymes of alanine transaminase, aspartate transaminase, and alkaline phosphatase were higher in the micro-encapsulated probiotic group compared to the two groups of probiotics and alginate with chitosan without the presence of bacteria, and all the three groups performed better compared to the control (P<0.05).CONCLUSIONS: Microencapsulation of probiotics maximizes the efficiency of probiotics in reducing oxidative stress.

BACKGROUND: There is growing evidence that resistance to antibiotics is increasing, and because of their wide range of negative impacts on the environment and humans, also accumulative effects in fish meat and hazards for humans, there have been numerous studies conducted using medicinal plants as a healthier alternative to antibiotics.OBJECTIVES: This study examined the antibacterial effect of two native medicinal herbs (oregano and echinacea mixed essential oil) on Lactococcus garvieae compared with some commercial antibiotics used in fish farms.METHODS: The antibacterial activities against a fish bacterial pathogen (Lactococcus garvieae) were tested using the disk diffusion method, followed by the determination of minimum inhibitory (MIC) and bactericidal (MBC) concentrations via sequential dilution of essential oil by Broth Micro Dilution method.RESULTS: The results showed antimicrobial activities of mentioned mixed essential oil with MIC ≥ 6.25 μL/mL and MBC ≥ 12.5 μL/mL. Also, the measurements of the diameter of inhibitory zones are consistent with the results of MIC and MBC, indicating the positive effect of this plant mix oil Lactococcus garvieae. However, the Lactococcus garvieae was more sensitive to Enrofloxacin (NFX 5), and Sulfamethoxazole &amp; Trimethoprim (SXT) (P<0.05), but the combination of Oregano and Echinacea essential oils performed better than Florfenicol (FF) despite the weaker effects against mentioned two antibiotics.CONCLUSIONS: The results approved the antibacterial activity of the mixed essential oil of oregano and Echinacea against Lactococcus garvieae on the laboratory scale. Using these compounds as a possible substitute for antibiotic compounds requires more clinical studies.

BACKGROUND: Bovine Herpes Virus-1 (BHV-1) belongs to the  Alpha herpesviral family. The virus is the cause of Infectious Bovine Rhinotracheitis (IBR) and Bovine Abortion. In the initial infection, the virus proliferates excessively. Moreover, shedding the virus leads to conditions in the latent phase of the disease. Infectious Bovine Vulvovaginit (IPV ) is the genital form of the disease that represents a genital infection and transmits via pustules and mucopurulent secretions. Exposure to the virus in genital mucosa leads to IPV infection through mating or artificial insemination and the diseases that can be transmitted to healthy livestock by frozen sperm during artificial insemination.OBJECTIVES: Viral contamination of the semen is one of the routes to spread the disease among dairy cattle. Therefore, we investigated the presence of the virus in domestic and frozen imported semen consumed in industrial dairy cattle farms.METHODS: In the present study, 140 frozen straws were collected. After melting each straw, 200 µl of obtained semen was used for DNA extraction, which was done directly on the semen samples and via a Genome Extraction Kit. Subsequently, to ensure the accuracy of the extraction, the PCR technique was done using PRM-1 gene primer. Tracking the viral genome was done using the PCR technique and known primers.RESULTS: In total, one out of 140 samples was found to be virally contaminated, and IBR contamination was confirmed by repeating all the steps and determining the gene sequence.CONCLUSIONS: It is necessary to further investigate the possibility that contamination can be transmitted via frozen semen, given that even one out of 140 samples is contaminated, and the importance of the disease.

BACKGROUND: Intensive aquaculture and commercialization production are believed to be needed for improving fish health status and probiotics are candidates to achieve this goal. OBJECTIVES: In this study, we utilized Lactobacillus plantarum and Lactobacillus pentosus as autochthonous food supplementation for biochemical investigation in rainbow trout. METHODS: In the current study, 240 juvenile rainbow trout with an average weight of 15 ± 2 g were randomly divided into four groups with three replications. Group 1, 2, and 3 were fed with 108 CFU/g of L. plantarum, L. pentosus, and the combination of these two probiotics, respectively. The control group was fed with the normal diet, for 60 days (2 % of body weight, 3times per day). Samples were taken at days 0 and 60 and biochemical parameters and hepatic enzymes activity were then investigated. RESULTS: Most biochemical factors had positive changes in the fish fed with probiotics, particularly in L. plantarum group (p < 0.05). Furthermore, the results exhibited a significant decrease in ALP level in the probiotic groups, particularly in L. pentosus group (p < 0.05). Meanwhile, AST enzyme activity in plantarum+pentosus group was higher than that in the other groups (p < 0.05). CONCLUSIONS: The present results indicated the effectiveness of food supplementation with probiotics in fish diet with the preference of probiotic to improve the biochemical factors and hepatic enzymes activity of rainbow trout, Oncorhynchus mykiss.

BACKGROUND: Backyard poultry are at risk of exposure to various viral contagious diseases such as Newcastle (ND) and Avian Influenza (AI). These diseases, in addition to the backyard poultry infection have an influence on villagers’ livelihoods. Also, backyard poultry plays an important role in circulation and survival of these viruses in environment and are considered as a risk factor for the poultry industry. OBJECTIVES: Studying the prevalence level of ND and Influenza H9N2 diseases in backyard chickens in Iran, in 2014-2015. METHODS: A descriptive cross-sectional study was conducted for two years (2014-2015) in backyard chickens with mortalities suspected of infection with ND or AI H9N2 viruses. Each mortality report was considered as one outbreak. For detection of possible ND or influenza virus infection tracheal and lung tissue samples were investigated by RT-PCR reaction test. Results were analyzed statistically by SPSS software. RESULTS: Overall, 121 outbreaks of Newcastle or influenza (H9N2) disease with 25.936 cases of death from 17 provinces were reported in two years. of  these, 54 outbreaks (44/6 %) were caused by H9N2 influenza virus, 58 (47/9 %) by velogenic ND virus and 9 (7/4 %) outbreaks were caused by influenza and Newcastle concurrent infection. Hotspot ratio in 2015 was significantly higher than in 2014. In comparison with Newcastle disease alone or concurrent ND and influenza outbreaks, the highest mean mortality rate was observed in H9N2 outbreaks. Outbreaks were reported in all seasons but the rate of occurrence in the months of June and July was significantly higher than the rest of the year. CONCLUSIONS: According to our results ND and H9N2 influenza virus infections are widely distributed in backyard chickens of villages in Iran. So, for implementation of control strategies, education of villagers, vaccination and annual surveillance of backyard poultry seem necessary.

BACKGROUND: It is possible for Sex reversal of eggs with female embryo using anti-aromatase agents, and the hatchlings have an improved performance. OBJECTIVES: The present study was conducted to survey the effect of in ovo injection of 0.1 mL Nettle (Urtica dioica) and mushroom extracts and Fadrozole hydrochloride as anti-aromatase agents  on sex differentiation of broiler chickens. METHODS: 366 fertile eggs from Ross 308 strain broiler breeders were divided into 4 groups. At the beginning of the fifth day of incubation in ovo injection operation was carried out. A control group was injected with 0.1 mL distilled water. The hatched chickens were then reared in a completely randomized design with 4 treatments and 6 replicates for 42 days. RESULTS: In ovo injection of 0.1 mL of fadrozole hydrochloride, mushroom and nettle extracts led to 100, 66.67 and 37.5 percent sex reversal, respectively (p < /em><0.05). During the whole of the experimental period, daily feed intake and daily weight gain of chickens of fadrozole hydrochloride and mushroom extract groups were higher than those on the control and nettle extract (p < /em><0.05). Fadrozole hydrochloride reduced levels of serum triglyceride and VLDL (p < /em> <0.05), but there was no significant difference in cholesterol, HDL-cholesterol and LDL cholesterol levels (p < /em>> 0.05). White blood cells and hematocrit percentage were not affected by experimental treatments (p < /em>> 0.05). Compared with other treatments, in ovo injection of mushroom extract caused a significant increase in the number and average diameter of muscle fibers (p < /em><0.05). CONCLUSIONS: The findings of this study confirm the significant anti-aramatogenic properties of the mushroom extract and the positive effect of in ovo injection of  mushroom extract on the sexes reversal and improvement in performance of broiler chickens.

BACKGROUND: Bovine tuberculosis caused by Mycobacterium bovis is an important disease that has negative effects on public health and entails economic loss. Traditional controlling programs for cattle  focus on test and slaughter strategy, and false positive is one of the disadvantages associated with tuberculin skin test. To overcome this limitation, proteins with high specificity have to be utilized. OBJECTIVES: The objective of this study was to clone and express virulent protein CFP-10 from Mycobacterium bovis AN5. METHODS: Full-length genes of cfp-10 were amplified by PCR technique. In parallel, pET23a(+) and PCR products were double digested by EcoRI and HindIII. Ligation was performed at 16˚C followed by transformation into competence E. coli DH5α. After being identified with sequencing, the cloned vector was transformed into E. coli BL21. Induction was performed by isopropyl-β-D-thiogalactopyranoside (IPTG). Urea 8M was used to dissolve the expressed protein in the inclusion body form. Recombinant protein was purified by Nickle-Resin, and urea was eliminated by decreasing the gradient. RESULTS: The CFP-10 gene clone was proved by sequencing method. The CFP-10, as a 10 KDa protein, was confirmed by Western blotting using monoclonal antibodies. Based on the results, the recombinant protein was successfully cloned and expressed. CONCLUSIONS: The results showed that cfp-10 gene was successfully cloned and expressed in prokaryotic system, indicating that this recombinant protein could be utilized in diagnostic kits against bovine tuberculosis in the future.

BACKGROUND: Various flea species have already been reported from dogs, among which the most important ones include Ct. felis, Ct. canis, and P. irritans. Fleas can cause annoyance in dogs and human and transmit a variety of bacterial, fungal, and viral agents to the host. In addition, they could function as an intermediate host of Dipylidium caninum and Hymenolepis diminuta.  OBJECTIVES: Due to the lack of molecular species-associated identification data, we conducted the current study to differentiate Ct. felis and Ct. canis with molecular assay. METHODS: In the present study, 605 fleas were primarily collected from the dogs referred to Tehran Veterinary Faculty hospital. Subsequently, the flea species were identified under a microscope with morphological keys. Afterwards, COX1 genes of Ct. felis and Ct. canis were amplified via PCR and the locus was finally compared utilizing RFLP and sequencing. RESULTS: Totally, 605 fleas were isolated from 20 dogs. In morphological studies, three species were identified: Ctenocephalides felis, Ctenocephalides canis, Pulex irritans. Pulex irritans had the highest frequency (61.8 %). In molecular study, 552 bp fragment of COX1 gene in two species was amplified and seen on agarose gel. After sequencing, it was seen that two species sequences in COX1 locus had a similarity of 99 % and all of them depended on Ct. canis. In PCR-RFLP, in which Taq1 enzyme was used for differentiation of two species, the same result was obtained. CONCLUSIONS: Even though these two species of dog flea are distinct morphologically, their molecular differentiation using COX1 genes was not successful.

BACKGROUND: It is fundamental to find the safest food preservatives in order to acheive the maximum health and economic benefits. Salmonella enteritidis is the cause of Salmonellosis, one of the most important foodborn zoonoses in poultry. OBJECTIVES: The present study was conducted to comparatively evaluate the antimicrobial effects of Trachyspermum copticum, Thymus vulgaris, and Cinnamomum zeylanicum essential oils and Formaldehyde against Salmonella Enteritidis. METHODS: In this study minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Cinnamomum zeylanicum, Trachyspermum copticum, and Thymus vulgaris essential oils against Salmonella enteritidis were determined using broth macro-dilution. The effects of essential oils on growth curve of Salmonella Enteritidis were also evaluated. RESULTS: In this study, MIC for the minimum inhibitory concentration level of Trachyspermum copticum, Thymus vulgaris, Cinnamomum zeylanicum and formaldehyde for Salmonella Enteritidis was determined as 500, 700, 500, and 70 ppm while MBC was calculated as 700, 900, 1000, and 200ppm, respectively. This study revealed that all the treatments increased lag phase. Bacterial growth speed was slower for Trachyspermum copticum of all the concentrations, yet a significant difference was observed only in 1 and 2 MIC of other treatments (p < /em><0.05). CONCLUSIONS: In conclusion, Trachyspermum copticum (with less concentration), Thymus vulgaris, and Cinnamomum zeylanicum (with higher concentration) could be employed as a substitution of a proporation of formaldehyde to prevent bacterial development in food.

BACKGROUND: Microbial growth and unfavorable taste and aroma are the leading reasons of meat spoilage under refrigerated temperatures. Today, plant essential oils are good alternatives to synthetic preservatives. In the meantime, nanotechnology has been instrumental in helping to apply the most effective plant compounds on the target cell. OBJECTIVES: The present study aimed to identify the compounds of essential oil of Cuminum cyminum and to investigate the microbial counts (total count, psychrotrophs, lactic acid bacteria, mold and yeast) after preparation of nanoemulsions of cumin essential oil via ultrasonic method and its nanoemulsions. Moreover, herein, we intended to evaluate organoleptic in chicken fillet samples under temperature of 4 °C. METHODS: The essential oil components were analyzed with GC / MS and the effects of Cuminum cyminum essential oil nanoemulsion were examined on microbial count and organoleptic properties in terms of color, odor, and overall acceptance (with 9-point hedonic scale) on Cuminum cyminum EO nanoemulsions in chicken fillets under 4 °C. RESULTS: The major identified components of Cuminum cyminum by extraction time were: beta- pinene (8.90 %), benzene- methyl (16.51 %), gamma- terpene (13.39 %), propanal (29.43 %), 1 -Isopropylidene-3-N-butyl2-cyclobutene (6.52 %), and benzene methanol (8.36 %). The results of microbial count at the last day of storage showed that the treatment containing 20 % and 25 % nanoemulsions significantly delayed microbial spoilage compared to other treatments (p < /em><0.05). The control treatment also indicated the highest level of microbial spoilage; as represented in Log 6, the maximum bacterial count in chicken fillets and the shelf life of BHT at 0.02 % concentration continued until day 3. Furthermore, in organoleptic evaluation, 20 % and 25 % nanoemulsion treatment scored higher than other treatments. CONCLUSIONS: Nanoemulsions of Cuminum cyminum EO at concentrations of 20 % and 25 % were able to delay the microbial spoilage of chicken fillets and produced organoleptically favorable results.