مجله تحقیقات دامپزشکی
سال هفتاد و ششم شماره 2 (تابستان 1400)

Cystic echinococosis (CE) is known to be one of the most important zoonotic diseases in different parts of Iran. Even though it causes major health problems, there is limited information regarding its transmission cycles and strain of this infection in eastern Iran.

The present study aimed to characterize the strain of Echinococcus granulosus cysts in the slaughtered sheep and goats in Birjand area using morphological and molecular criteria.

Isolates of E. granulosus were collected from sheep (30) and goats (30) from Birjand slaughterhouse and characterized employing both DNA (PCR-RFLP of ITS1) and morphological criteria (metacestode rostellar hook dimensions). In addition, the fragments of the genes coding for ITS-1 were sequenced.

Among the two different identified strains/genotypes (sheep and camel), the sheep strain appeared to be the most common genotype of E. granulosus affecting sheep and goats. All of the 30 sheep samples and 20 out of 30 goat samples were infected with sheep strain. However, the camel genotype was only observed in the goats and 10 out of 30 goat isolates were infected with the camel genotype. The camel genotypes had RFLP patterns, which were different from the RFLP patterns of the rest of isolates (sheep strain).

The results of this study revealed that the ‘sheep’ strain was the most prevalent strain in sheep and goats in this area. Moreover, the camel genotype (G6) was confirmed to trigger infection in the slaughtered goats of Birjand area.

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. 

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.

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.

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.

Even though these two species of dog flea are distinct morphologically, their molecular differentiation using COX1 genes was not successful.

Toxocaiasis is an important zoonotic disease caused by the second stage larvae of Toxocara canis and Toxocara cati. Toxocariasis is the most common worm infection in several temperate countries and causes severe complications. C-type lectin is one of the larval stage products of this parasite. It is involved in immune responses, including cellular signals in vertebrate immunity, activation of innate immunity in vertebrates and non-vertebrates, and induction of homeostasis. 

The current research aimed to optimize the production of recombinant C-type lectin of Toxocara canis and investigate its antigenic properties.

Reference nucleotide sequence of lectin type C (CTL) of Toxocara canis (T. canis) was extracted from Genbank. Recombinant Plasmid, pET32a, including the desired sequence was then constructed by GENERAY. The recombinant plasmid was transformed to Escherichia coli strain BL21 (DE3). The expression of the recombinant protein was investigated using SDS-PAGE and dot blot methods and approved with human T. canis positive serum.

The findings of the present study showed that optimization and high level production of recombinant protein expression was achieved by selecting Escherichia coli (BL21 (DE3), 37 °C temperature for 4 hours after induction and 1 mM IPTG concentration. After optimization, the recombinant protein was obtained at a concentration of 1160±0.6 µg/mL. The molecular weight of the resulting recombinant protein was 42 kDa. Recombinant plasmid passage in Escherichia coli DH5α strain caused a significant increase in recombinant protein expression. The results of condition optimization evaluation, with SDS-PAGE and Dot blot methods, showed that the highest production of recombinant C type lectin protein of T. canis was obtained under optimized conditions.

Based on these results, to produce reasonable amounts of specific C-type lectin recombinant protein, further studies are needed to evaluate its immunogenicity and protection against Toxocara canis infection.

Full supply of the nutritional requirements of C57BL/6 mice plays an important role in preventing ulcerative dermatitis syndrome.

We conducted this work to asses ulcerative dermatitis in C57BL/6 mice colony and reduce it through dietary modification.

During a two-years period, a C57BL/6 mice colony in a laboratory animal breeding center was monitored for the presence of ulcerative dermatitis syndrome. In the first year, the animals were fed with outbred laboratory mice pellet and in the second year, the modified diet was used based on the nutritional requirements of this inbred mouse.

The average numbers of litters per parturition were 3 and 6 in the first year and the second year, respectively. The litter's mortality rate in the first year was 75 % and zero in the second year. The total number of mice remaining in the colony was 220 in the first year, which increased to 1100 in the next year. Syndrome symptoms were observed in the adult mice in the first year. Alopecia was observed mainly in the back and neck and the severity of the symptoms was mild to severe. No mortality was observed in the mice, even in severe cases, and their recovery was spontaneous, yet very slow. In the necropsy, axillary lymphadenopathy, splenomegaly, and hepatomegaly were observed. In histopathological samples taken from the skin lesions, the accumulation of inflammatory cells and the spongiosis of cornea cells were observed. In lymph nodes, spleen, and liver, pathological changes were observed from the accumulation of inflammatory cells to necrosis. In the second year, the number of cases reduced significantly to 2 %. The severity of the symptoms was mild and their recovery was faster than that in the previous year.

By fully supplying the nutritional requirements of C57BL/6 mice, we could significantly prevent the occurrence of ulcerative dermatitis syndrome.

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.

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.

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.

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).

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.

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.

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.

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.

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.

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.

Medicinal plants have a long history in the treatment of parasitic diseases and are usually free of side effects. Removing and killing tomonts and theronts of this parasite can prevent the parasite pathogenicity in fish.

The present study aimed to investigate the inhibitory effect of ethanolic extract of Zataria multiflora on both free and parasitic life stages.

Three soluble forms of Zataria multiflora extract (80 ml/L) were prepared, which contained freshly prepared solution, stored solution at room temperature for one month and stored solution at refrigerator for one month. Afterwards, different dilutions were prepared for each of the solutions and the tomonts and Theronts were treated with different dilutions of the extract.

Zataria multiflora extract can kill all theronts at concentrations of 20 ml/L during 6.04 to 6.37 min and concentration of 10 ml/L during 31/2 to 32.4 min, respectively. However, at these concentrations, only 3.46-6.11 % of tomonts were killed herein after 4 hours.

Tomonts were found to be more resistant to the parasiticides than theronts. The use of Zataria multiflora extract significantly reduced tomont reproduction and decreased Ich infectivity prevalence and intensity. Zataria multiflora extract prevented Ich infestation in naive fish and effectively treated infected fish.

Abomasal ulcer is a disease of digestive system in ruminants. The clinical signs vary widely depending on the type of abomasal ulcer. The economic losses of this disease include decreased milk yield and mortality.

The present study aimed to determine the frequency, type of ulcer, pathological, hematological, and electrocardiographic indices in the slaughtered cattle in Sanandaj abattoir.

A total of 400 cattle were randomly selected with both sexes and 1-2, 2-3, 3-4, and ≥4 years of age. They were grouped during three seasons and the abomasal ulcer was evaluated. Blood sample was collected and electrocardiogram was recorded before they were slaughtered. Abomasal lesions were assessed macroscopically and histopathologically in the slaughtered cattle.

Out of the 400 abomasom examined, 51 (12.75 %) were affected by abomasal ulcers, all of which were type-1 ulcer. Subtypes 1a, 1b, 1c, and 1d. 35 cattle (68.7 %) had the types of arrhythmia. The most frequent of them was sinus arrhythmia and sinus tachycardia. Neutrophilia and reduction in MCHC were significantly more than the other hematological indices (p < /em>˂0.05). There was a significant relationship between the prevalence of abomasal ulcers and arrhythmia, gender, age, and season (p < /em>˂0.05). Furthermore, there was a significant relationship between electrocardiographic and hematological indices in the affected cattle (p < /em>˂0.05).

The results of this study revealed a high prevalence of abomasal ulcer in the studied area. They also indicated that the electrocardiographic and hematological changes could be helpful in the diagnosis of abomasal ulcer.

Magnetic cell targeting is a novel non-invasive cellular delivery technique. It improves stem cell delivery to and retention in the injury site. Labeling cells with superparamagnetic iron oxide nanoparticle (SPION) is one of the most important steps of this technique. Appropriate SPIONs selection is believed to be of vital importance.

The current study aimed to produce SPIONs which are capable of attaching to Mesenchymal stem cells surface (MSCs).

Dextran coated SPIONs were produced following co-precipitation method under N2 atmosphere. Bone marrow derived MSCs were isolated and cultured from rabbit humerus bone. Anti-rabbit CD44 monoclonal antibody was attached to the surface of SPIONs and MSCs and were labeled with this final product. SPIONs coating process, particle size, and antibody conjugation efficacy were evaluated using FT-IR, SEM, and Bradford protein measurement assay, respectively. Attachment of antibody-linked dextran coated SPIONs to MSCs was accessed utilizing Prussian blue staining, immunofluorescence analysis, and SEM analysis.

Peaks of FT-IR at 3200 cm-1 and 2922 cm-1 are representative of dextran. The average particle size was 56.13±6.67. The average antibody-SPION conjugation ratio was 77.78±6.35%. The average percentage of the labeled cells in Prussian blue and IF analysis were 71.57±2.53 and 95.04±0.95, respectively. MSCs-SPIONs conjugation was also confirmed via SEM analysis.

In conclusion, it could be inferred that mesenchymal stem cells could successfully be labeled with dextran coated-anti CD44 antibody conjugated- superparamagnetic Iron oxide nanoparticles. This product could be used for further in-vitro and in-vivo evaluations.

The regulation of appetite and food intake in birds are implemented as complex homeostatic mechanisms at different levels of control.

The current research aimed to investigate the effects of glycine and strychnine-sensitive receptors on food intake induced with dopamine in neonatal broiler-type chickens.

This study was conducted in five experiments (each consisting of four treatment groups with 12 birds). In experiment 1, chickens in the control group received intracerebroventricular (ICV) injection of saline (with 0.1 % Evans Blue) and different doses of dopamine (10, 20, and 40 nmol) in treatments groups 2-4, respectively. Experiments 2 and 3 were designed similar to the experiment 1 except for the fact that chickens received different doses of glycine (50, 100, and 200 nmol) and strychnine (50, 100, and 200 nmol) instead of dopamine. Experiment 4 was performed to investigate the mediatory role of strychnine (100 nmol) on food intake induced with dopamine (40 nmol). Moreover, experiment 5 investigated the interaction between non-effective doses of glycine (50 nmol) and dopamine (10 nmol) and their interplay on food intake. Afterwards, cumulative food intake based on bodyweight percentage (BW %) was measured at 30, 60, and 120 minutes after the injection.

The obtained findings revealed that effective doses of dopamine and glycine dose-dependently induced hypophagia in neonatal meat-type chickens (p < /em>≤0.05). In addition, injection of strychnine increased food intake and also inhibited the hypophagic effect induced by dopamine (p < /em>≤0.05). Furthermore, co-administration of non-effective doses of glycine and dopamine significantly decreased food intake compared to the groups which only received dopamine or glycine (p < /em>≤0.05).

Our findings suggested that strychnine-sensitive receptors may have a mediatory role in food intake induced by dopamine. Additionally, it seems that glycine and dopamine probably have synergistic effects on food intake control in neonatal meat-type chickens.

Oxidative stress is involved in pathogenesis of Polycystic Ovary Syndrome (PCOS). Lutein is a herbal compounds with antioxidant properties.

The current research aimed to evaluate the effect of lutein on body weight and histomorphometry of uterus in experimental PCOS induced with Dehydroepiandrosterone (DHEA) in mouse models.

Twenty-four female NMRI mice aged 20 days and weighing 14-17 g were randomly assigned to four equal groups: control, experimental PCOS, and PCOS treated groups with 125 and 250 mg/kg lutein. The induction period of PCOS with oral administration of DHEA (6 mg/100 g, daily) was 21 days and lutein treatment was followed by the induction period of 28 days. The mean body weight of the groups was evaluated on day 0, day 21 (at the end of DHEA treatment), and day 49 (at the end of treatment period) with lutein. The mean diameter of the uterine wall, the mean overall thickness of the uterine wall, the average thickness of the endometrium, myometrium and uterine epithelium, along with the number of endometrial gland branches were measured utilizing light microscope.

The results revealed that body weight in the PCOS group was significantly higher than that in the control group on days 21 and 49. Treatment with 125 and 250 mg/kg of lutein reduced body weight in the lutein treated groups compared with PCOS (p < /em><0.01). The mean uterine wall diameter, mean total uterine wall thickness, mean thickness of endometrium, myometrium, and uterine epithelium with the number of uterine endometrial branching were significantly lower in the control and lutein treated groups compared to those in the PCOS group (p < /em><0.05). The use of both doses of lutein (125 and 250 mg / kg) significantly improved uterine histopathological indices, particularly the mean uterine wall diameter (p < /em>=0.0001) compared to the PCOS group.

Lutein could improve the side effects of induced PCOS by DHEA on body weight and uterine parameters.

Several studies have reported the application of mesenchymal stem cells (MSCs) for the treatment of equine musculoskeletal disorders. Bone marrow (BM) and adipose tissue (AT) are the main sources for isolation of MSCs. Although the MSCs derived from various sources have similarities, certain differences have been reported in terms of biological, proliferative, immunological, and differentiative characteristics. They are also different concerning gene expression profiles. So, comparative gene expression analysis on mRNA level via quantitative real-time polymerase chain reaction (qRT-PCR) is believed to be of great value for better characterization of MSCs from different sources.

Mastitis could be detected in various ways, including physical, on-farm, and laboratory tests.

The present research aimed to evaluate the accuracy of the diagnosis of mastitis using lactate dehydrogenase (LDH)-based dipsticks and to assess these dipsticks with regard to the effects of different lactation days, the amount of milk production, and parity.

Considering bacteriologic culture as a gold standard method for the diagnosis of subclinical mastitis, the sensitivity and specificity of LDH test were 68.9 % and 54 %, respectively. The results revealed a high correlation between SCC and LDH. A statistically significant relationship was observed between the response of dipstick and CMT results; with the increase in the CMT score, the score of LDH dipstick increased. By investigating the effects of lactation days, the amount of milk production, and parity, it was determined that the chance of having subclinical mastitis in cows with positive dipstick result was 5.59 times greater than that in cows with negative dipstick result. There were no significant relationships among SCC, LDH, and CMT with lactation days and milk production; meanwhile, with the increase in parity, the three above-mentioned variables showed significant increase.

The results of the present study indicated that the best methods for subclinical mastitis detection were SCC, CMT, and LDH based dipsticks, respectively.

Clostridium perfringens is an important animal pathogen that causes severe loses to the livestock and poultry industries. Therefore, bacterial detection is believed to be of particular importance.

The present study aimed to identify Iranian isolates using conventional and molecular methods and to evaluate their toxicity.

In this work, 23 Clostridium perfringens type B isolates were examined via microbiological and biochemical tests. Subsequently, they were subjected to PCR technique for the final confirmation. After culturing of the isolates in specific medium, the minimum lethal dose test was performed. The most toxigenic isolate and reference strain was prepared the enterotoxaemia anaculture vaccine. Serum neutralization test was performed on the experimental inactive vaccines.

The results revealed that etx and cpb gene could be found in all of the isolates, yet cpb2 gene was found in 65.2 % of the isolates. The minimum lethal dose ranges for these bacteria was less than 1/10 to more than 1/900. The results of serum neutralization in Iranian isolate and reference strains were 5 and 10 IU / ml, respectively.

The findings herein implied that strain 1795 with high toxicity could be used in vaccine production. Of course, for use in production, further research on target animals is needed.

HISTORY:

Suspensory ligament injury associated with avulsion or crescent fracture of third metacarpal or metatarsal bone is a common injury in racehorses. Lameness is known to be the most common clinical sign of these horses.

CLINICAL FINDING:

The present article investigated two avulsion fractures in the third metacarpal bone in 2.5-year-old mare and stallion, which had chronic lameness with a vague origin and disability of weight bearing in left forelimbs. DIAGNOSTIC TESTS: Following radiography and ultrasonography examinations, suspensory ligament injury and subsequently avulsion fracture in the proximal third of metacarpal bones in left forelimb were approved.

FINAL EVALUATION:

In radiography, radiolucent lines associated with fragmented segments (about 2×6×8 mm) in mare and (about 2×4×6 mm) in stallion  in the proximal part of  the third metacarpal bone were indicative of an avulsion or crescent fracture at the origin of the suspensory ligament. Furthermore, in ultrasonography compatible with radiographic findings, an echogenic-fragmented fracture in the proximal part of the third metacarpal bone and a local area of decreased echogenicity was diagnosed as desmopathy of suspensory ligament.