Veterinary Research Forum
Volume:10 Issue: 3, Summer 2019

Analysis of elements content in honey is important for honey quality and safety and for monitoring of environmental pollution. The levels of 22 elements, aluminum (Al), barium (Ba), calcium (Ca), cadmium (Cd), chromium (Cr), cobalt (Co), iron (Fe), copper (Cu), potassium (K), magnesium (Mg), manganese (Mn), nickel (Ni), sodium (Na), lead (Pb), strontium (Sr), silver (Ag), bismuth (Bi), gallium (Ga), indium (In), lithium (Li), thallium (Tl) and zinc (Zn), were determined in 70 samples obtained from beekeepers located in the West Mediterranean region of Turkey. Determination of elements content was carried out using ICP-OES. Chromium, Co, Cd, Ag, Bi, In and Tl were not detected in any of the tested honey samples. The most abundant metal was K which has an overall average of 764.26 mg kg-1. Higher concentrations of Pb, Ni, Mg, Na, K and Mn were found in the samples obtained from Burdur compared to other provinces. The levels of Cu were statistically lower in Antalya in comparison to other regions. No significant differences were observed in Al, Zn, Fe, Sr, Ba, Ca and Ga levels between regions. The differences in the chemistry of honey samples collected from different regions may be due to geochemical soil composition and geographical differences. Their levels were below to the European limits and the honeys are safe for human consumption.

Acetamiprid (ACP) belonging to the neonicotinoid family used against wide array of pests in agriculture and domestic purposes. In this study, we evaluated the attenuating effects of ethanolic extract of Mangifera indica leaves (EEMI) in averting reproductive toxicity caused by ACP in male guinea pigs. Thirty male guinea pigs were randomly assigned to five treatment groups (n = 6). Group 1 (T0) received distilled water orally; group 2 (T0-) was given 80 mg kg-1 of ACP and groups 3, 4 and 5 were treated, respectively, with EEMI at doses of 50, 100 and 200 mg kg-1 plus ACP. After 90 days, the reaction time, sexual organ weights, sperm count, motility and anomalies, spermatozoa with entire plasma membrane, testicular histology, serum testosterone concentration, testicular malondialdehyde (MDA) level, reduced glutathione (GSH) concentration, testicular superoxide dismutase (SOD) and catalase (CAT) activities were assessed. Co-administration of EEMI significantly reduced the reaction time, sperm anomalies and testicular MDA, SOD and CAT levels compared to the T0- group. Co-treatment of EEMI significantly alleviated sperm count and motility, percentage of spermatozoa with the normal plasma membrane, serum testosterone concentration, accessory sex gland weights, and testicular GSH concentrations. The ACP treatment induced cell membrane degradation in the testis and this effect was prevented with the addition of EEMI. In conclusion, ACP negatively affected the animal reproductive function and induced oxidative stress. The addition of EEMI alleviated the toxic effects of ACP on the reproductive function of male guinea pigs.

In the present study, the effect of different concentrations of cell-free supernatant (CFS; 10.00 and 35.00 mg g-1) of Lactobacillus salivarius (Ls-BU2) on chemical, microbial and sensorial specifications of ground beef stored under the refrigerated condition was investigated. The antibacterial activity of CFS on Escherichia coli was also assessed. According to agar-disk diffusion method, CFS of Ls-BU2 revealed a promising antibacterial activity against E. coli in culture media compared to CFS of a well-known probiotic (L. acidophilus LA-5). In meat, CFS of Ls-BU2 showed a minimal effective concentration (MEC) of 35.00 mg g-1 on E. coli, while CFS of L. acidophilus represented a MEC of > 45.00 mg g-1. The CFS of Ls-BU2 at 35.00 mg g-1 concentration retained psychrophilic counts of meat at a lower value than maximum accepted level (7 log10 CFU g-1). In a similar trend, CFS of Ls-BU2 at 35.00 mg g-1 concentration was also displayed high sensorial scores compared to other CFS-treated samples. In conclusion, we demonstrated that CFS of Ls-BU2 and to some extent CFS of L. acidophilus could act as a safe food additive for the control of bacterial pathogens and to extend the shelf life of ground beef.

Neuro-immune mediators play an important role in the development of sickness behaviors. In the present study, the effect of captopril on sickness behaviors caused by lipopolysaccharide (LPS) was studied in the rats. The animals were randomized into the following groups: control, sham, 10 mg kg-1 captopril - LPS (Capto 10-LPS), 50 mg kg-1 captopril - LPS (Capto 50-LPS), and 100 mg kg-1 captopril - LPS (Capto 100-LPS). Behavioral tests including open-field (OF), elevated plus maze (EPM) and forced swimming (FS) test were performed, and the serum level of interleukin-6 (IL-6) was assessed. In OF, the number of crossings in the central zone in Capto 10-LPS, Capto 50-LPS, and Capto 100-LPS groups was higher than that of the sham group. In EPM, the open arm entry numbers in the sham group were lower compared to the control group. Furthermore, pretreatment by captopril increased the entries to the open arms. In FS test, the immobility time of the sham group was longer than that of the control group. In Capto 10-LPS, Capto 50-LPS, and Capto 100-LPS groups, immobility was shorter compared to the sham group. In addition, the IL-6 level was higher in the sham group compared to the control group, and treatment with 50 and 100 mg kg-1 of captopril restored the IL-6 level in comparison with the sham group. Results confirmed that pretreatment with captopril ameliorated LPS-caused sickness behaviors and attenuated IL-6 as an inflammatory marker in the rats.

Polyclonal antibodies against kappa light chain are used to diagnose diseases producing free light chain. The kappa and lambda light chains are products of immunoglobulin synthesis and released into the circulation in minor amounts such as serum, cerebrospinal fluid, urine and synovial fluid in normal condition. The purpose of this study was the production and purification of polyclonal immunoglobulin G (IgG) against human kappa light chains. In this study, early human IgG was purified by ion-exchange chromatography, reduced with Dithiothreitol and heavy and light chains were separated with size-exclusion chromatography. Afterward, affinity chromatography with protein L Sepharose at pH 2.00 was displayed to be a dominant condition for the separation and purification of the kappa light chain of immunoglobulins from human serum. Eventually, the rabbit was immunized by human kappa light chains. The rabbit IgG was purified and labeled with horseradish peroxidase (HRP). Direct enzyme-linked immunosorbent assay was planned to determine the titer of HRP conjugated rabbit IgG against the human kappa light chain. The optimum titer of anti-kappa IgG was 1:16000. At the result, purified polyclonal anti-kappa is useful tool in biomedical and biochemical researches and diagnostic kits.

Chick embryo extract (CEE) contains a variety of growth factors which may improve in vitro follicle growth. Therefore, the effect of CEE on mouse pre-antral follicle culture was evaluated. Different percentages of CEE (0, 0.50%, 1.00%, 5.00% and 10.00%) were added to culture medium. Hence, the osmolarity of media was measured. Pre-antral follicles with diameter of 120-150 μm were isolated from 12-14 days old mouse ovary and cultured for 12 days. After culture, the maturation rate was assessed. Granulosa cells viability was evaluated using MTT test and estradiol levels were evaluated using related radio-immunoassay (RIA). Genes expression (BMP15 and ALK6) was also evaluated. The osmolarity of media and granulosa cells viability were the same in all groups. Estradiol level in group with 10.00% CEE was significantly decreased compared to the control group. After 12 days culture, the percentage of antral follicles development was significantly higher in the group with 5.00% CEE compared to control group. The percentage of metaphase II and germinal vesicle breakdown oocytes was significantly higher in group 5.00% CEE compared to control group. The expression of BMP15 gene in antral follicles in 5.00% CEE and control groups was significantly lower compared to pre-antral follicles. However, the expression of ALK6 gene in antral follicles in 5.00% CEE and control groups was not significantly different compared to pre-antral follicles. The increasing effect of CEE on follicle viability with keeping normal gene expression indicates that addition of proper percentage of CEE to culture media improves culture conditions, making it a possible choice to be used as a follicular growth enhancer in infertility clinics.

Canine seminal plasma contains antioxidant enzymes to protect sperm against internally generated ROS. These enzymes are removed from seminal plasma during the process of cryopreservation. The freezing/thawing process can cause some morphological and functional changes via ice crystallization and osmolality imbalance. The present study was conducted to evaluate the effects of curcumin supplementation on sperm total count, motility, progressive motility, viability, morphology, total antioxidant capacity (TAC), DNA integrityand NOX5 gene expression of dog frozen semen. The pooled semen was allocated to fresh (Group 1) and frozen (Group 2) controls, curcumin (2.50 mM) (Group 3) and curcumin (5.00 mM), (Group 4). Sperm parameters including total sperm count, morphology, motility, progressive motility, sperm concentration and DNA integrity in addition to TAC were evaluated in fresh and frozen-thawed semen samples. Real-time PCR was used to investigate NOX5 and GADPH (reference gene) genes expressions. Curcumin at 2.50 mM provided a greater protective effect on the DNA integrity compared to 5.00 mM and control groups. TAC was significantly higher in 2.50 mM group than other groups. NOX5 gene expression in curcumin 2.50 mM was higher than 5.00 mM group. In conclusion, curcumin seems to emolliate sperm parameters and to protect sperm against sperm reactive oxygen stress and increases NOX5 gene expression.

Crocin, as a carotenoid compound of saffron, exerts a potent antioxidant property. Mesalazine is frequently used in the treatment of ulcerative colitis. This study investigated the effects of separated and combination treatments with crocin and mesalazine in a rat model of ulcerative colitis. Ulcerative colitis was induced by intra-colonic administration of acetic acid (4.00%, 1.00 mL) at 8 cm proximal of the anus. Normal saline, acetic acid, crocin (5.00, 10.00 and 20.00 mg kg-1), mesalazine (100 and 300 mg kg-1) and crocin (5.00 mg kg-1) plus mesalazine (100 mg kg-1) were administered after induction of colitis for eight days. Body weight, oraganosomatic index (OSI), macroscopic and microscopic evaluations of colon and measurement of malondialdehyde (MDA), superoxide dismutase (SOD), tumor necrosis factor-alpha (TNF-α) contents of colon tissue were determined on day eight after induction of colitis. Crocin (10.00 and 20.00 mg kg-1), mesalazine (300 mg kg-1) and crocin (5.00 mg kg-1) plus mesalazine (100 mg kg-1) significantly (p < 0.05) improved body weight and OSI and reduced macroscopic and microscopic scores. These treatments also significantly (p <0.05) recovered the increased levels of MDA and TNF-α as well as the decreased level of SOD in colon tissue. Crocin and mesalazine did not produce significant effects in intact rats. Based on the results, it is concluded that crocin and mesalazine produced protective effects on colon tissue via antioxidant and anti-inflammatory actions. In addition, a synergistic effect was observed between crocin and mesalazine in attenuating ulcerative colitis.

Natural compounds including essential oils (EOs) are used to inhibit the pathogenic bacterial growth in foods. The objective of this study was to investigate the combined effect of Trachyspermum ammi EO and propolis ethanolic extract (PEE) on some food pathogenic bacteria. In this study, the effect of T. ammi EO and PEE at different concentrations, individually and in combination, on Escherichia coli, Salmonella typhimurium, Listeria monocytogenes, Bacillus cereus, and Staphylococcus aureus were investigated using minimum inhibitory concentration and fractional inhibitory concentration (FIC) indices. Moreover, the effect of sub-inhibitory concentrations of EO and extract on E. coli and S. aureus growth curve was determined. The results revealed the synergistic effect of T. ammi EO and PEE on S. aureus, B. cereus, E. coli, S. typhimurium, and L. monocytogenes. The effects of EO and extract sub-inhibitory concentrations on bacterial growth curve resulted in an increased lag phase of the culture and decelerated bacterial growth, especially S. aureus. The results showed that T. ammi EO and PEE, especially when combined, are effective in bacterial growth inhibition and their potential use in food systems is recommended.

The aim of this experiment was to investigate rumen fermentation and some blood parameters of dromedary camels fed with C3 and C4 forage. Four fistulated dromedary adult camels were fed with diets as a changeover design, 30 days for each period. The diets included alfalfa hay + wheat straw (C3 forage) and atriplex+ suaeda + seidlitzia (C4 forage). At the end of the experiment, rumen and blood parameters, gas production of wheat straw and atriplex as a 2 × 2 factorial experiment were determined. The highest blood glucose and urea nitrogen levels were found for camels fed with C3 forage, 2 hr after feeding (p < 0.05). The maximum NH3-N concentration in the rumen was for diets C3 and C4, 2 and 4 hr after feeding (p < 0.05). The lowest rumen pH was observed for C3 diet at 2 and 4 hr and for C4 diet at 4 and 8 hr after feeding. The activity of rumen carboxymethylcellulase (CMCase) and microcrystalline cellulase (MCCase) enzymes was the highest for C3 and C4 diets, 8 hr after feeding, however, during feeding the enzyme activity in C4 was higher than that of 2 hr (p < 0.05). The rumen volatile fatty acid (VFA) concentrations were significantly higher in camels fed C3 forage in comparison with C4 (p < 0.05). The results showed that the gas production potential was significantly higher in treatments containing atriplex, however, the gas production rate was higher in treatment containing wheat straw (p < 0.05). The results suggested that for camels maintained in closed systems, the replacement of C3 forages instead of C4 could be possible and useful.

Influenza is a highly contagious zoonotic disease in the world. Avian H9N2 influenza virus is a significant pandemic pathogen widely distributed throughout the world. Pet ownership has been documented as a risk factor for infection transmission to human. Considering major public health concern, the prevalence of antibodies against avian H9N2 influenza virus was evaluated in 170 serum samples of dogs by hemagglutination inhibition assay. This study is the first survey to assess the epidemiology of avian H9N2 influenza virus infection in dogs in Kerman, southeast of Iran. Out of 170 samples, 65 (38.23%) were positive for H9N2. Antibodies were higher in farm dogs that were kept with other animals and also in dogs were fed a raw diet. These findings emphasize the importance of close attention to these populations for control and prevention programs. It is important to reduce infection burden, especially in regions with widespread distribution of H9N2.

This experiment was conducted to investigate the effects of olive leaf (OL) on the performance, abdominal fat pad and some ileal bacterial population of Cobb broiler chickens. A total number of 400 day-old chicks were randomly distributed into floor pens and reared under the same condition until 14 days of age. On day 14, each pen was randomly assigned to one of the five experimental treatments with four replicates of 20 male and female chicks. The dietary treatments were consisted of a control group which fed basal diet without OL entire period of the study and groups 2 to 5 that fed diets supplemented with 0.25, 0.50, 0.75 and 1.00% OL powder, respectively. On days 21 and 42 of the experiment, ileal digesta samples were collected under the sterile condition to evaluate ileal bacterial population. The results indicated that birds fed diets containing various levels of OL, had higher body weight gain (except for 1.00% OL) and lower feed conversion ratio compared to that of the control group. Dietary inclusion of OL resulted in a higher count of Lactobacillus sp.  compared to the control group on 42 days of age, while Escherichia coli count significantly was not influenced. The abdominal fat pad was lower in birds fed OL supplemented diets. In conclusion, findings of the current experiment showed that the OL had positive effects on feed conversion ratio, abdominal fat pad deposition and ileal bacterial count of broiler chickens.

Immune-mediated hemolytic anemia (IMHA) is a common cause of anemia in dogs. The immune aggression towards erythrocytes can be triggered by many pathological conditions such as infection, inflammatory disease or neoplasia. Upon ruling out any eliciting conditions, a diagnosis of the primary immune-mediated disease can be made. In this particular case of severe anemia (tested positive for circulating antibodies against red blood cells with flow cytometry), vector-borne diseases (which are a common cause of immunopathology in Mediterranean countries) were excluded, leptospirosis was not. This resulted in an unsuccessful immunosuppressive therapy with prednisone, two whole blood transfusions and ultimately death of the patient. Leptospirosis (confirmed positive in two tests, micro-agglutination test for antibodies and PCR for microbial DNA in urine), can mimic a primary IMHA and must be considered in its differential list of causes. A liver involvement, that included elevated serum activity of liver enzymes and increased serum bile acid was observed at the admission and suggested an etiopathogenesis other than a primary IMHA.

After sudden death with a history of about two weeks ruminal tympany, a 3-year-old, male alpaca from Huantaiqi Zoo, Chongqing, China was presented to the Animal Diseases Rapid Diagnosis Center, Southwest University, Chongqing, China for diagnosis of the death causes. At necropsy, the primary pathological lesions were found in the lung. A pronounced hemorrhage with topical congestion and lobular pneumonia was identified. Sero-fibrinogenous pleural effusion was also detected in the thoracic cavity. After necropsy, the lung sample was processed for histological examination, while lung, hydropericardium, and heart-blood samples were processed for bacteriological examination. From the lung tissue, abundant fluid exudate was found in the pulmonary alveoli. Meanwhile, a mild to moderate hemorrhage and inflammatory cells infiltrations were also observed in the lung sections. Pure isolates on the 5.00% defibrinated sheep blood agar were submitted for identification by morphological and molecular methods. Sequencing results indicated that the Gram-negative sporadic bacilli were all belonged to Morganella morganii. To the best of our knowledge, this is the first record of M. morganii induced pneumonia in an alpaca.