Iranian Journal of Veterinary Medicine
Volume:11 Issue: 4, Autumn 2017

Packaging is one of the important aspect of food quality and safety. Unfortunately, most of food packaging materials are from oil resources that are limited resources and undegradable.
The aim of this study was reaching an environmentally friendly packaging with enhancement safety of food.
After obtaining Bunium persicum (BP) and Mentha pepperita (MP) essential oils (EOs) by steam distillation method, their chemical compositions were determined using GC-MS analysis. PLA films were prepared using solvent casting technique containing different concentrations of BP (0, 0.5 and1%v/v), MP (0, 0.5 and1%v/v) EOs and cellulose nanoparticle (CN) (0 and 1% w/v) and their antimicrobial effects against gram positive bacteria (Staphylococcus aureus ATCC 65138, Bacillus cereus ATCC 11778) and Gram negative bacteria (Vibrio parahaemolyticus ATCC 43996, Escherichia coli O157:H7 and Salmonella typhymurium ATCC 14028) were assessed by disk diffusion method.
Major compounds of BP EO were Propanal 2-methyl-3-phenyl (34.08%), Cymene (18.23%) and Myrtenal (12.37%) and for MP EO were p-Menthan-3-ol (44.59%) and p-Menthan-3-onetrans (12.14%). The results of present study indicated that pure PLA film or PLA films containing CN showed no antimicrobial activity against any of the five tested bacteria but films containing EOs had significant antimicrobial activity and BP EO was more effective than MP EO and their combination (p The results of this study indicated that PLA films containing MP and BP EOs may be useful for packaging of foods in order to increase their shelf life and safety.

Bacterial resistance to antibiotics is a crucial public health problem. Essential oils (EOs) possess antimicrobial effects and have been screened as potential natural antimicrobial compounds. This study was conducted to compare the effects of Eugenia caryophyllus (clove) and Origanum compactum (oregano) EOs on the growth of Staphylococcus aureus and the expression of the SEA, SEC and SEE genes.
The minimum inhibitory concentrations (MIC) of EOs and growth of bacterium at subMIC levels of EOs was determined. Enterotoxin detection was done using a commercial SE visual immunoassay kit after 18, 24, 48 and 72 h. Gene expression of enterotoxins was evaluated through RNA extraction, DNA synthesis and performing real time-PCR using specific primers for each SE.
MIC of clove and oregano were 2 µl/ml and 1µ l/ml, respectively. Colony counts at 48 and 72h of cultures grown at 75% MIC of clove oil showed the growth rate was reduced 1.67 and 1.83 log10 cfu/ml compared to the control, and in the case of oregano at 75% MIC the decreases in growth rate were 2.25 and 2.68 log10 cfu/ml, respectively. When the target bacterium is cultured in the presence 75% subMIC of EOs, the transcript levels of sea, sec, see and the regulatory gene (agrA) were decreased 8.81, 9.13, 9.08 and 8.32 fold in the case of clove, and 11.56, 9.96, 11.07 and 11.15 fold in the case of oregano, compared to the control.
The growth, gene expression and as a result secretion of enterotoxins A, C and E by S. aureus were decreased significantly at subMIC levels of EOs, especially at 75% MIC.

One of the major aspects of traceability in food authenticity assessment is to explore practical methods to find the origin of food.
The aim of the present study was to find a DNA based method for authentication and traceability of food, which are of great importance in health management.
Four different DNA extraction methods were applied to obtain high pure DNA in some oil samples including olive oil, sunflower, canola and soybean oil to improve the traceability. The isolated DNA was analyzed by PCR using common primer pair, derived from the region harboring 18S rRNA/5.8S rRNA genes. Extraction methods were developed based on specific binding of DNA molecules to the silica membrane (column) or resin.
Our results showed that amplifiable DNA could only be extracted from olive oil in method 1, whereas the isolated DNA from other samples needed to be purified. In method 2, by pre-treating of oil with PBS and subsequent precipitation with Isopropanol, the amplification of isolated DNA was observed in sunflower, crude canola and olive oil. To remove more effectively the contaminants, method 2 was combined with chloroform and resin/Isoporopanol precipitation as method 3. Interestingly, the extracted DNA from all examined oil samples could be amplified with mentioned primers. For elimination the disadvantages of chloroform, method 4 was set up by direct usage of lysis and binding buffer. The extracted DNA from all refined oil samples could be amplified successfully.
Based on our findings, the major problem in DNA extraction from oils is the PCR inhibitors in extracted DNA, which can be resolved by the presented methods 3 and 4.

Phosphorous is one of the expensive nutrients in poultry feed. Therefore, improving the bioavailability of this nutrient in feed ingredients could be effective for lowering the cost of feed.
This study was performed to evaluate the effect of pre-treatment of feed ingredients by commercial enzymes and different levels of pH on releasing of phosphorus from phytate under in vitro condition.
Three solutions including Distilled water, HCl 0.5% and HCl 1% (with pH=5.5, 2.12 and 2.12 respectively) and three enzymes (None, Bio-phytase, Rovabio Excel AP, and Rovabio Max AP) were used to determine phytate content of corn and soybean meal. First, each sample was supplemented with the enzymes and pre-treated under above mentioned solutions for 3 hours at 25 and 40ºC.
The results indicated that pre-treating of corn samples with Bio-phytase or Rovabio Max AP and different solutions (at 25 ºC and 40 ºC for 3 hours) reduced phytate content significantly. The best results obtained with corn samples supplemented with Rovabio Max AP and mixing by HCl 1% at 25ºC, so that phytate content decreased up to 99.5% in comparison with control. The same results were also obtained for soybean samples. The highest reduction in phytate content (up to 47.4%) was observed by adding Rovabio Max AP and HCl 1% solution at 40ºC.
It could be concluded that pre-treating of corn and soybean meal with several manners such as using commercial enzymes including phytase and solutions with different pH were effective to reduce phytate content that means increasing bioavailability of phosphorous.

Considering widely administration of methylphenidate and also its immunosuppressive effects on different organs, importance of related microscopic studies is obvious.
Determining histological effects of methylphenidate on adrenal glands and lymphatic organs in mice.
A total number of 30 adult male Balb/C mice were provided, weighed and divided into one control and two experimental groups. The control group received water by gavages once a day, for 40 days. The experimental groups were orally administered MPH hydrochloride (2mg/kg and 10mg/kg body weight,) respectively. Animals were anesthetized and blood samples were collected through cardiac puncture for analysis of blood cells. Spleen, thymus, lymph nodes and adrenal glands were removed and processed for microscopic studies through hematoxylin and eosin staining. Spleen samples were processed for plasma cell count and staining (label antibody CD138*). The data were analyzed using analysis of variance (ANOVA) and p

The growth and differentiation of skeletal pectoral limb girdle, wing and the ossification centers in these regions after hatching were investigated in some groups of quails.
The aim of this study was to determine the age of physical maturity and radiological and histological assessment of the ossification centers of pectoral limb in quail.
14 quails after hatching were reared in similar and standard conditions and sampled once every 7 days to 90 days.
According to radiological and histological results, differentiation of the wing in quail commences with the appearance of centers of undeveloped cartilages in diaphyseal humerus, radius, and ulna at the end of 7 days, and also carpal regions at the beginning of the 14 days. The growth sequence in humerus, radius, ulna, carpus, metacarpus, and digits are observed in various stages that the high growth is related to the maximum cartilaginous activity and their ossification stages and humerus keeps its growth connection constant with the length of the whole wing skeletal, although its growth scale lessens after 21th day. The histological results were evaluated based on prepared tissue sample from the proximal humeral portion. Lack of bone marrow was observed in the all 1th day`s tissue samples and bone marrow conformation was commenced after 7th day. The growth plate was not observed in the all samples and this issue is complementary to the information obtained from radiographic examination.
According to this study, complete the ossification process and the formation of all parts of pectoral limb girdle and wings in the quails is 70 days after hatching.

In order to analyze the cecum-colon ecosystem and the treatment of the cecal impaction and hindgut acidosis, cecal cannulation is needed. It is essential to select a simple, fast and inexpensive cecal cannulation method. Because of different complications in general anesthesia, the standing surgery is known as a better option for the horse emergency surgery.
The objective of the present study was to design a simple, fast and inexpensive cecal cannulation method in standing horses.For the standing surgery, a new approach was designed and experimented on four horses.
For this purpose, at first a cannula with approximately 7cm, 2cm and 2.6cm in length, internal and external diameters, respectively was designed. Immediately before the standing surgery, the horses were sedated with xylazine (1mg/kg) and morphine (0.3 mg/kg). After incising the subcutaneous tissue, the external abdominal oblique, internal abdominal oblique and transverse abdominis muscles were opened by grid incision. The peritoneum was bluntly perforated and the abdomen was exposed. The muscles were separated only enough to permit one hand to enter the abdomen. The cecum was readily identified by palpation of the cecal base and the dorsoventrally oriented tenia. At this stage, a purse string was secured on the serosal surface of the cecum by nylon and a stab incision was made. Then the cannula was inserted into the cecum and the suture was tightened.
The surgery was successfully performed for all horses, however, some complications such as increasing body temperature, transient signs of colic, ileus, pneumoperitoneum, subcutaneous emphysema and necrosis of the borders of the skin in the sutural places were detected. All complications were alleviated by proper nursing management.
The surgical method was successfully terminated. Therefore, the method is recommended as a simple and inexpensive emergency surgical method for cecum in order to conduct different investigation, diagnosis and treatment techniques.

Azathioprine is an immunosuppressive agent that is used in a variety of dermatologic, digestive and hematologic disorders in both humans and small animals.
Aims: The effects of long term Azathioprine administration on complete blood count, rate of CD4 and CD8 lymphocytes, serum immunoglobulins and protein concentrations of mixed-breed dogs were evaluated in this study.
24 healthy mixed-breed dogs were divided randomly into two equal control and treatment groups. Dogs in the treatment group received the therapeutic dose of Azathioprine for four months, while the dogs in the control group didnt receive this drug. Peripheral blood samples were taken from both two groups before and after the trial to check CBC, CD4 and CD8 lymphocytes and the concentrations of total protein, albumin, serum IgM and IgG.
There was significant decrease in the levels of WBC, RBC, hematocrit and CD4 lymphocyte and double positive CD4/CD8 rates (all p values Myelotoxicity induced by Azathioprine could be the probable cause of decrements in the rate of WBC and RBC. Decrease in the rate of dpCD4/CD8 might be due to decrement in dpCD4/CD8 progenitor cells and/or decrease in the activation rate of single positive T cells as the result of pharmacological effect of Azathioprine. Disrupted synthesis processes, from genes to proteins through Azathioprine might be the cause of decreases in the level of serum gamma globulins and protein.

Semen cryopreservation might be influenced by sperm concentration. This study was conducted to investigate three different semen concentrations [100 (C100), 50 (C50), and 25 (C25) × 106 spermatozoa/mL] on freezability of bull semen.
On each collection day, four ejaculates were collected (a total of 24 ejaculates from four bulls), pooled and divided to three equal parts. Each part was diluted to reach to one of the above mentioned final semen concentration and then frozen. After thawing, sperm motility, apoptosis status and mitochondrial activity were assessed.
The results showed that C100 resulted in significantly higher total sperm motility compared to C50 and C25 groups. The percentage of live spermatozoa was significantly higher in C100 compared to C50 and C25 groups. Also, C25 resulted in significantly higher early and late apoptotic spermatozoa compared to C50 and C100 groups. The mitochondrial activity was significantly lower in C25 compared to C100 and C50 groups.
It seems that low sperm concentrations (as low as 50×106) may be less advantageous for cryopreservation.

The immunostimulating effect of Aloe vera in mammals has been documented, but few works were done on effect of A. vera on fish health and immune responses.
In this study the effect of oral administration of A. vera on growth indices, hematological parameters and immune responses of rainbow trout were investigated.
One thousand five hundred rainbow trout fingerling (20 ± 2 g, Mean ± SD) were divided into five groups, each in triplicate, in farm scale. Group 1 were adopted as control and fed with non-supplemented feed, groups 2 to 5 were fed with diet supplemented by 0.05%, 0.1%, 0.2% and 0.5% A. vera extract respectivly for 60 days. Growth indices (SGR, FCR, PWG, FER, PER and CF) calculated in day 30 and 60. Blood samples were taken in day 60 and hematological parameters including: PCV, Hb, RBC, WBC, MCH, MCV, MCHC as well as immunological parameters including: Lysozyme and serum bactericidal activity, serum total protein and globulin were compared among the groups.
Results showed that all calculated growth indices (except CF) and all mentioned immunological parameters were significantly increased in fish fed with 0.1% and 0.2% A. vera supplemented food (G3 and G4) compare to control group (P 0.05).
It can be concluded that oral administration of 0.1% and 0. 2% A. vera crud extract in food (G3 and G4) can improved growth indices, stimulate non-specific immune responces and affect some hematological parameters positively in rainbow trout.