Veterinary Research Forum

Abstract This study aimed to determine the effects of in-feed inclusion of humic substances (HS) and/or calcium carbonate (CaCO₃) on the performance and welfare of laying hens. A total number of 144 Hy-Line Brown laying hens (55 weeks old) were randomly assigned to four treatments, T1–T4 (36 birds per treatment). T1 hens were fed on a control diet without HS or CaCO₃, T2 hens were fed on the control diet + 2.00 g per bird per day CaCO₃, T3 hens were fed on the control diet + 0.20% HS and T4 hens were fed on the control diet + 0.20% HS + 2.00 g/bird/day CaCO₃. The experiment started after 15 days of adaptation and lasted 8 weeks. The parameters evaluated were percentage of hen-day egg production, food consumption, mortality, egg quality parameters and heart rate variability (welfare indicator). Hens in the T3 group showed a significantly lower feed intake than those in the other three groups, however, significantly higher daily egg production was recorded in groups T3 and T4 compared to T1 and T2. Eggshell quality characteristics were significantly improved by HS supplementation and eggs laid by hens in groups T1, and T4 presented paler yolk and shell color than those in the other groups. In conclusion, these results indicated that in-feed inclusion of HS had a beneficial effect on laying hens’ productive performance including egg production and eggshells quality.

Abstract The purpose of this study was to investigate the tetracycline resistance in Trueperella pyogenes isolates from bovine samples in Burdur, Turkiye, and assess 16 tetracycline-resistance genes distribution among the isolates. Forty-nine T. pyogenes isolates were phenotypically characterized for anti-microbial resistance to doxycycline, oxytetracycline and tetracycline by disc diffusion method. Presence of tetracycline genes of T. pyogenes was investigated by multiplex and singleplex polymerase chain reaction. Our results indicated that 87.80% and 42.86% of the isolates were resistant to tetracycline and oxytetracycline, respectively, and the rate of resistance to doxycycline was 6.12%. Total of 21 (42.85%) were carrying tetracycline-resistance genes and tet(A) was present in 12 (24.49%) isolates; whereas, the tet(W) gene was identified in 9 (18.37%) and 2 (4.08%) of the isolates carried both tet(A) and tet(W), respectively. The study indicated antibiotic resistance patterns of tetracycline agents and links to the tet-genes among T. pyogenes were detected. It makes it worthwhile that this is the first report for detection of tet(A) gene in T. pyogenes.

Abstract Despite being important, there are no equations for prediction of ionized calcium (iCa) in sheep and cattle. The objectives of this study were i) to create equations for the calculation of serum iCa concentration based on the serum concentrations of total calcium (tCa), albumin (Alb) and total proteins (TP) and ii) to investigate whether predicted serum iCa values are beneficial in clinical practice. Serum samples from 30 sheep and 30 dairy cattle were used. Serum tCa was determined colorimetrically, while serum iCa was determined with an ion selective electrode method. Serum Alb and TP concentration were determined using bromo-cresol green and biuret methods, respectively. Ionized calcium was also calculated based on serum tCa, using regression analysis, and with two equations based on Alb and TP concentration. Bland–Altman plots were plotted to evaluate the agreement between measured and predicted iCa; Passing and Bablok (P - B) regression analysis was used to assess their agreement. The initial equations were corrected using the P - B generated equation and Bland – Altman plots were run to evaluate the level of agreement between measured and predicted iCa using the final equations. Six equations were finally created for cattle and 6 for sheep. The total bias exceeded 10.00% in all of them indicating that they are clinically unacceptable for iCa prediction especially when the predicted result is very close to the cut-off point of < 1.00 mmol L^-1. So, it could be suggested that, when necessary, iCa concentration should be directly determined.

Abstract The emergence and spread of multidrug resistance among pathogens of the agro-food sector is increasing at an alarming rate, which has directed attention to the search for alternative to antibiotic therapy. The present work studied the physiological and population dynamics of lytic bacteriophages against avian-adapted Salmonella. Out of 28 positive samples, four bacteriophage isolates (SalØ-ABF37, SalØ-RCMPF12, SalØ-MCOH26, SalØ-DNLS42) were selected based on their ability to clearly lyse bacterial test strains. The isolates propagated were active against closely related D1 serotypes, i.e., S. Enteritidis and S. Typhimurium, with no heterologous activity against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 23235. Each of the monophage suspension and cocktail efficiently suppressed the bacterial count from exponential increase in comparison to the untreated bacterial control. The bacterial turbidity was recorded as 0.244 at λ₆₀₀ during 400 min of co-incubation, in contrast to bacterial control showing λ₆₀₀ = 0.669. The latent period was recorded to be 25, 35, 25 and 30 for SalØ-ABF37, SalØ-RCMPF12, SalØ-MCOH26 and SalØ-DNLS42, with 73.00, 97.00, 132 and 75.00 PFU cell^-1, respectively. The highest lytic activity was seen at 37.00 ˚C - 42.00 ˚C, with phage particle count being fairly stable at pH 3.00 - 9.00. Each of the isolates possessed dsDNA by being resistant to RNase A. The current study concludes that lytic phages are promising alternative to combat multidrug resistant superbugs. The physiological characterization and bacterial growth inhibition are important parameters in standardization of phage therapy.

Abstract Diabetes mellitus is one of the leading causes of death globally. The development of cellular injuries and impaired energy metabolism are involved in the pathogenesis of diabetes mellitus, leading to severe diabetic complications in different tissues such as the pulmonary tissue. Autophagy is a double-edged sword mechanism required for maintaining cell survival and homeostasis. Any abnormalities in autophagic response can lead to the progression of several diseases. Here, we aimed to assess the effect of diabetic conditions on the autophagic response and exosome secretion in a rat model of type 2 diabetes mellitus. The experimental diabetic group received 45.00 mg kg^-1 streptozocin (STZ) dissolved in 0.10 M sodium citrate. After 4 weeks, we monitored autophagic response and exosome biogenesis in the pulmonary tract using immunohistochemistry (IHC) and Real-time polymerase chain reaction analyses, respectively. Histological examination revealed the interstitial bronchopneumonia indicating enhanced immune cell infiltration into the pulmonary parenchyma. Immunohistochemistry staining displayed an enhanced autophagic response through the induction of microtuble-associated protein light chain 3 (LC3) and protein sequestosome 1 (P62) compared to the control rats. These changes coincided with significant induction of tetraspanin CD63 in STZ-induced diabetic rats relative to control rats. In conclusion, a diabetic condition can increase the autophagic response in pulmonary tissue. The accumulation of P62 in the pulmonary niche exhibits an incomplete autophagic response. The abnormal autophagy response can increase pulmonary cell sensitivity against injuries.

Abstract The aim of the present study was to evaluate the efficacy of macrolides to eliminate intramammary infection (IMI) caused by Staphylococcus aureus (S. aureus) and Streptoccus spp. 3 weeks before calving time. Eighty Holstein dairy cows with subclinical mastitis pathogens were divided into three groups. Three weeks before expected parturition time, cows in group 1 received tilmicosin (n = 29), cows in group 2 received tylosin (n = 30) and cows in group 3 were left as negative control (n = 21). Milk samples were obtained on 3 and 7 days after calving. Randomly amplified polymorphic DNA (RAPD) method was determined for all of the S. aureus isolates that had the same isolates before and after parturition. The total cure rate was 63.33, 75.86 and 66.66% for tylosin, tilmicosin and control groups, respectively. Furthermore, cure rates were not significant, when each type of mastitis causing pathogens were considered separately. The incidence of clinical mastitis during 60 days after calving for tylosin, tilmicosin and Control groups was 23.33, 27.58 and 38.09%, respectively. Only four S. aureus isolated before drying-off were similar to post-calving isolate, according to RAPD-PCR method. In conclusion, antibiotic therapy before calving improved the cure rate numerically, however, it was not significant.

Abstract Among the 26 human claudin proteins, the food-poisoning bacterium Clostridium perfringens produces an enterotoxin (~ 35.00 kDa) that specifically targets human claudin 4, causing diarrhea by fluid accumulation in the intestinal cavity. The Clostridium perfringens enterotoxin (CPE) C-terminal domain (cCPE ~ 15.00 kDa) tightly binds to claudin 4 and disrupts the tight junction barriers in the intestines. In this study, we aimed to determine the contribution and type of amino acid interactions involved in association between claudin 4 and the C-terminal CPE. First, the three-dimensional format of claudin 4 was downloaded from RCSB. Then, during 60.00 nanoseconds (nsec), molecular dynamics simulation was conducted using the GROMACS package on CPE of crystallographic structure. The results indicated that the simulations performed well during the simulation times and there were no noticeable problems or artifacts. We found that Coulombic (glycine 317, proline 311 and serine 313) and Lennard-Jones (tyrosine 310, leucine 315, serine 313 and glycine 317) interactions played a significant role in complex stability. This information localized the C-terminal of CPE as a linear sequence sufficient for recognition and binding to the eukaryotic CPE receptor. A detailed description of the dissociation process brings valuable insight into the interaction of the claudin 4-cCPE_290-319 complexes, which could help in the future to design more potent drugs.

Abstract In the recent years, the use of medicinal plants to reduce the effects of mycotoxins in foods and feeds has been considered. This study was conducted to investigate the effects of Aloe vera on performance, serum biochemical parameters and liver histopathology in laying hens fed on aflatoxin B1 (AFB1)-contaminated diet. Seventy-two White Leghorns (Hy-Line W-36) were randomly allocated to four treatments. 1) basal diet (control), 2) control plus 1.00 mg kg^-1 AFB1, 3) control diet plus 1.00 mg kg^-1 AFB1 + 100 ppm Aloe vera powder, and 4) control diet plus 1.00 mg kg^-1 AFB1 + 300 ppm Aloe vera powder. Each treatment consisted of three replicates of 6 birds. Egg weight and Haugh units were not affected by AFB1. Egg production and eggshell thickness were lower for groups fed 1.00 mg kg^-1 AFB1. Egg production, egg weight and eggshell thickness were improved by incorporation of Aleo vera in the AFB1 contaminated feed but were not significant. Chickens fed AFB1 had significantly lower aspartate aminotransferase (AST), alanine aminotransferase (ALT) and uric acid and higher cholesterol than other groups. Aloe vera powder improved levels of cholesterol, uric acid, AST, and ALT. AFB1 also caused histopathological changes in liver tissues, such as vacuolar degeneration, fatty infiltration, and necrosis. The addition of Aloe vera powder to the aflatoxin containing diet reduced the severity of lesions in liver. The data demonstrated the ability of Aloe vera to reduce the adverse effects of AFB1 exposure in laying hens.

Abstract Polycystic ovarian syndrome (PCOS) is a complex endocrine and metabolic disorder. Chlorogenic acid (CGA) bears antioxidant properties with protective effects on different tissues. This study was conducted to evaluate the effect of CGA on follicular development, hormonal status and biomarkers of oxidative stress in a rat model of PCOS. In this experimental study, 18 rats were divided into three equal groups including: control, non-treated PCOS [(estradiol valerate (EV): 40.00 mg kg^-1 intramuscularly)], and PCOS-CGA (EV: 40.00 mg kg^-1 intramuscularly and CGA: 100 mg kg^-1 intraperitoneally once a week for eight consecutive weeks). At the end of treatment period, all rats were anesthetized. Then 5.00 mL blood samples of rats in the three groups were taken and prepared for hormonal analyses and their ovaries were isolated and dissected mechanically free of fat and mesentery. The ovaries underwent the following analyses: Morphological study with Hematoxylin and Eosin staining and biochemical study using the malondialdehyde (MDA) level and total antioxidant activity. Data were analyzed using one-way ANOVA and post hoc Tukey’s test. The serum level of luteinizing hormone, estrogen, testosterone, antioxidant capacity, glutathione and the number of cystic follicles in the PCOS group treated with 100 mg kg^-1 Chlorogenic acid compared to the non-treated PCOS group were significantly decreased, however, the serum level of follicle stimulating hormone, progesterone, MDA and the number of secondary, graafian follicles and corpus luteum were significantly increased. Chlorogenic acid could be effective in ameliorating follicular development as well as hormonal and biochemical disorders in rats with PCOS.

Abstract Stage X is one of the formation stages in birds at which the blastoderm area is distinguished by two areas of area pellucida being responsible for formation of embryonic tissues and primordial germ cells, and area opaca forming the extra-embryonic tissues. Primordial germ cells are multi-potent stem cells giving rise to spermatogonia or oogonia. The present study was carried out to describe the characteristics of primordial germ cells in stage X of pheasants’ embryo using a transmission electron microscope. The blastoderm was dissected out from embryos which were already incubated for 12 hr. Toluidine blue was used for staining semi-thin sections; lead citrate and uranyl acetate were also used to stain ultra-thin sections. Images of primordial germ cells elucidated that the nucleus was situated eccentrically and had a compact spherical structure. Moreover, the nucleolus appeared elongated and was located eccentrically. The cytoplasm was composed of yolk granules and glycogen particles. Mitochondria were observed as round structures in the cytoplasm. The most important finding was that the primordial germ cells contained yolk granules, mitochondria and small amount of glycogen at this stage.

Abstract Resistance to the knockdown effect of pyrethroid insecticides occurs due to mutations at target sites of pyrethroids, meaning the voltage-gated sodium channels gene (VGSC) in the membrane of the neurons. In fleas, this mutation occurs at two sites in the sodium channel in neurons: one is the replacement of leucine with phenylalanine (L1014F) and the other is the replacement of threonine with valine (T929V). In this study, 81 Pulex irritans and 47 Ctenocephalides canis fleas were collected from five provinces in the west and northwest of Iran. Adult fleas were exposed to cypermethrin 0.75%, and the mortality rate was calculated after 1 and 8 hr, and the mutation sites in the VGSC gene were investigated. The lethality of cypermethrin 0.75% for P. irritans was 40.00 - 57.14% after 1 hr and 60.00 - 73.91% after 8 hr. The lethality of this dose for C. canis after 1 and 8 hr of exposure was 33.33 - 41.17% and 66.66 - 80.33%, respectively. The VGSC sequence analysis indicated two mutation sites in the resistant and one mutation site in the susceptible fleas. The VGSC sequence analysis of susceptible P. irritans showed that 5.50% of them were homozygous susceptible and 94.45% were hetero-zygous susceptible. Susceptible C. canis were 5.26% homozygous and 94.73% heterozygous susceptible. All the resistant fleas were homozygous. The development of pyrethroid resistance and high-frequency L1014F mutation in fleas suggest that pyrethroids are likely to be ineffective in controlling fleas. Therefore, monitoring pyrethroid resistance and its underlying mechanisms are necessary for controlling fleas and finding new alternative control methods.

Abstract Due to the increasing demand for natural foods with fewer artificial additives, broccoli is considered by the consumers and researchers due to its taste, nutritional values and biochemical components. Therefore, this research was conducted to investigate the effect of broccoli juice and balsamic vinegar on the physicochemical and textural attributes of the beefsteak aimed at tenderizing it and decreasing the ageing period. Experiments were carried out through marinating the beefsteak with three different treatments containing 3.00 units protease g^-1 beefsteak of broccoli juice and 10.00% (v/w) of balsamic vinegar during 48 hr storage at 4.00˚C. A significant reduction in pH and Warner-Bratzler shear force as well as the increase in myofibrillar fragmentation index and myofibrillar protein solubility were obtained showing that the beefsteak was tendered in the presence of broccoli juice and broccoli juice+balsamic vinegar. v_

Abstract Expression and purification of human DT-diaphorase, also referred to as NAD(P)H quinone oxidoreductase 1 (NQO1; EC. 1.6.99.2), which is a flavoprotein belongs to the family of oxidoreductases are optimized. The DT-diaphorase plays an important role in biosensor design for laboratory analysis and also developing biosensor for measurement of glucose level in blood. The aim of this study was to investigate various parameters regarding the expression of DT-diaphorase in Escherichia coli BL₂₁ (DE3) and thermal stability of DT-diaphorase activity at different temperatures in the presence of sucrose. Expression conditions of DT-diaphorase in E. coli were optimized with an induction time (22.00 hr), induction temperature (18.00 ˚C) and also lactose (5.00 mM) and isopropyl ß-D-1-thiogalactopyranoside (1.00 mM) concentrations as inducers. The Km, Vmax and kcat values for NADH as a substrate were 25.50 µM, 357 µM per min and 446.40 μM mg^-1 per min, respectively. Results of our research revealed that different concentrations of sucrose at 40.00 ˚C did not have any significant effect on enzyme structure; while, relatively significant changes, especially in the presence of sucrose (0.75 M) at 50.00 ˚C were observed. The results presented show that sucrose causes DT-diaphorase inactivation rate reduction and relatively little increases in thermal stability and thus, sustains its conformation against thermal unfolding.

Abstract This study was carried out to investigate the effect of different hormonal programs on the reproductive parameters and serum progesterone concentration of Lake-Ghashghaei ewes in spring season. In this study, 144 ewes were selected from two large nomadic herds. The ewes were synchronized using progesterone sponges. One day before harvesting the sponges, all ewes were injected intramuscularly with 400 IU of eCG. Then ewes were divided into six groups (12 ewes). In the first herd, 1 (GnRH-1 group), 2 (GnRH-2 group), 5 (GnRH-5 group), 7 (GnRH-7 group) and 12 days (GnRH-12 group) after estrus observation, 25.00 µg GnRH was injected intramuscularly to the ewes. In the second herd, the ewes were received 500 IU of hCG on days 1 (hCG-1), 2 (hCG-2), 5 (hCG-5), 7 (hCG-7) and 12 (hCG-12). In each herd, a group of 12 ewes received only eCG and progesterone sponges and was considered as control group. The results showed that twinning rate, the number of born lambs, fecundity and prolificacy rate were higher GnRH and hCG received groups on the second day compared to other groups. The serum progesterone concentration was significantly higher in all groups received GnRH and hCG compared to control group. In general, the results showed that GnRH and hCG two days after mating improved different reproductive activities in addition to increasing serum progesterone concentration. On the other hand, hCG had a greater effect on progesterone concentration, the number of born lambs and the fecundity rate compared to GnRH.

Abstract Canine Distemper Virus (CDV) is the cause of a highly lethal infectious disease affecting a broad range of carnivores. Despite using various treatments, there is still no effective treatment, especially in the neurological form of distemper. The aim of this study was to evaluate the therapeutic effect of injecting Newcastle disease vaccine into the subarachnoid space of dogs with neurological form of distemper. The dogs that had symptoms of nervous distemper, particularly myoclonus, were included in the plan. After anesthetizing of dogs, 0.10 to 1.00 mL of cerebrospinal fluid (CSF) were removed and, 0.10 to 0.50 mL of the prepared Newcastle solution were injected into their subarachnoid space. Another 0.50 to 1.00 mL of normal saline was then injected to remove the needle from the vaccine. The live attenuated LaSota or B1 vaccine was used in this study. Rapid kit tests and reverse transcription polymerase chain reaction (RT-PCR) assays were used to diagnose of the disease. Dogs were monitored for up to 3 to 24 months during that time they were evaluated for improvement or worsening of clinical symptoms. Out of nine dogs in which distemper were diagnosed with different tests, one dog recovered completely and another dog recovered greatly. Therefore, the overall recovery rate was 22.20%. It is concluded that administration of Newcastle vaccine into the subarachnoid space of dogs with nervous distemper causes at least 22.20% improvement and does not cause specific side effects and can be used to treat affected dogs.

Abstract Avian pathogenic Escherichia coli (APEC) and uropathogenic E. coli (UPEC) can cause vast infections in humans and poultry. The present study was conducted to compare the isolates of the APEC and UPEC pathotypes on the basis phenotypic and genotypic features of antibiotic resistance and phylogenetic differences. Total number of 70 identified E. coli strains, including 35 APEC and 35 UPEC isolates, were isolated from avian colibacillosis and human urinary tract infection (UTI), and were subjected to the antimicrobial susceptibility testing, polymerase chain reaction (PCR) detection of the resistance genes, phylogenetic grouping and DNA fingerprinting with enterobacterial repetitive intergenic consensus PCR (ERIC - PCR) to survey the variability of the isolates. The most resistance rates among all E. coli isolates were, respectively, obtained for Ampicillin (84.20%) and sulfamethoxazole-trimethoprim (65.70%). The APEC and UPEC isolates showed the most susceptibility to imipenem and gentamycin, respectively. Among 70 APEC and UPEC isolates 34.20%, 32.80%, 20.00%, and 12.80% belonged to the A, B2, D, and B1 phylogenetic groups, respectively. Analysis of the DNA fingerprinting phylogenetic tree showed 10 specific clusters of APEC and UPEC isolates. According to the results, the most effective antibiotics and the phenotypic and genotypic predominant resistance patterns of the APEC and UPEC isolates were different. Moreover, APECs and UPECs showed various dominant phylogenetic groups. With all descriptions, the APEC isolates still are potential candidates for carrying important resistance genes and can be one of the possible strains related to human infections.

Abstract Chicken fillet is a suitable medium for growth and activity of different types of microorganisms. The pH and nutrients content of fillets are the most important factors in their microbial spoilage and degradation during cold storage at the retail level. In this regard, the uses of edible coatings containing antimicrobial and antioxidant compounds are effective approaches to maintain the quality of fillets. In this study the inhibitory effect of edible coating materials based on chia seed mucilage containing 0.00%, 0.25% and 0.50% zinc oxide nanoparticles (ZnO-NPs) on microbial growth and chemical spoilage as well as enhancing shelf life of chicken fillets during refrigerated storage for 20 days was investigated. The results of X-Ray diffraction confirmed the dispersion of ZnO-NPs on the chia seed mucilage matrix. Also, the number of total aerobic mesophilic and psychrophilic bacteria, coliforms and lactic acid bacteria, and the pH, total volatile nitrogen, peroxide and free fatty acids indexes in the control fillets were significantly increased compared to the fillets coated with chia seed mucilage during storage. While, in the samples coated with chia mucilage containing ZnO-NPs the number of the above-mentioned bacteria decreased in the first stage and then significantly increased during storing. Based on our findings, the shelf life of fillets could be increased at least 20 days by coating them with the chia seed mucilage containing ZnO-NPs.

Abstract Astigmatid mites are economically significant pests of stored products and sources of inhalant allergens causing allergic rhinitis and asthma worldwide. The morphological identification of astigmatid mites at the species level is often a difficult task due to their small size, phenotypic similarity and lack of diagnostic characters. We used multiplex polymerase chain reaction (PCR) to identify astigmatid mite species, which could complement the morphological data for the species-specific identification of mites. Internal ribosomal transcribed spacer (ITS) sequences (i.e., partial 18S, the full length of ITS1-5.8S-ITS2 and partial 28S) from eight astigmatid species (Acarus siro, Tyrophagus putrescentiae, Suidasia nesbitti, Dermatophagoides pteronyssinus, Dermatophagoides farinae, Lepidoglyphus destructor, Chortoglyphus arcuatus and Gohieria fuscus) were obtained by DNA extraction and then sequenced after PCR amplification. Specific primers were designed in the ITS2 region manually. Results revealed that an identification method for eight common astigmatid species was established based on multiplex PCR, which should be effective for the identification of other species of mites by redesigning species-specific primers in future experiments.

Abstract Epidermal growth factor (EGF) has biological roles, including embryonic organ development, breast morphogenesis, breast cell proliferation, and mammary development. This study aimed to measure EGF concentration and evaluate its relationship with somatic cell count (SCC) in healthy water buffaloes (Bubalus bubalis) milk. The study material was constituted of 120 milk samples obtained from 30 healthy water buffaloes between the ages of 3 - 6 years, negative for California mastitis test and SCC less than 3.00 × 10⁵ cells mL^-1 milk. In milk serum samples, the EGF concentration was measured using a bovine-specific enzyme-linked immunosorbent assay kit. Epidermal growth factor concentration in the buffalo milk was ranged from 4.30 to 9.80 ng mL^-1, with a mean of 8.30 ± 1.50 ng mL^-1. Positive correlation between milk SCC values and EGF concentrations was recorded in water buffaloes. Further research is required to evaluate the content of milk EGF in different species of animals because of the EGF effective role in mammary gland and intestinal mucosa.

Abstract The present study was conducted to investigate the detection and identification of Cryptosporidium species via molecular techniques and evaluate the serum concentrations of inflammatory factors in Cryptosporidium species. The fecal samples (n = 256) were collected from pre-weaned (≤ 2.00 months) calves and the positive samples were identified utilizing Ziehl-Neelsen staining. Nested species-specific multiplex PCR (nssm-PCR) and restriction fragment length polymorphism (RFLP) were used to identify the species and sub-species. The serum concentrations of IL-1β, IL-6, IL-12, TNF-α, and IFN-γ were also assessed. The results revealed that 10.54% of samples were positive. The results of Nested-PCR showed that 92.59% of the samples were positive for C. parvum while 7.41% were positive for C. andersoni. The results of RFLP confirmed 92.59% of the samples for C. parvum, 3.70% for C. muris / C. andersoni, and 3.70% for C. muris. The serum concentrations of IL-1β, IL-6, IL-12, TNF-α, and IFN-γ were significantly higher in the infected calves compared to those in healthy calves. However, the serum concentration of IFN-γ was significantly higher in the calves infected with C. parvum while the serum concentrations of TNF-α and IL-6 were significantly higher in those infected with C. andersoni. In conclusion, C. parvum was prevalent in the region and the calves demonstrated inflammatory responses to Cryptosporidium species.

Abstract No diagnostic kits and reagents are available in the market to detect and evaluate camel immune responses to different pathogens. This study aimed to produce sheep anti-camel (Camelus dromedarius) polyclonal antibodies (pAbs) and to determine the specificity with other species immunoglobulin. Immunoglobulins (Igs) from camel serum samples were purified using ammonium sulfate precipitation (40.00% saturated ammonium sulfate). Purity of the camel Igs was tested by sodium dodecyl sulfate polyacrylamide gel electrophoresis. PAbs against (Camelus dromedarius) immunoglobulins were generated by immunizing sheep with purified Igs. Anti- camel Ig polyclonal antibodies titer and specificity were determined using ELISA and Western blot techniques. Polyclonal antibodies specific to camel Igs were significantly high in immunized sheep which confirmed the immunization procedure. PAbs reacted specifically with camel serum immunoglobulin and did not react with other species immunoglobulin of horse and chickens. Polyclonal antibodies produced in this study can be regarded as a valuable tool to be used for immune-diagnostic purposes in camel population world-wide.

Abstract Diaphragmatic rupture is introduced as one of the most common injuries occurring in small animals. In our retrospective report, 14 cats with a mean age of 12 months (male = 8, female = 6) that underwent diaphragmatic herniorrhaphy were studied. The data included the following: age, sex, breed, and clinical signs. The cause of diaphragmatic hernia, diaphragmatic rupture area, herniated organs, concomitant injuries, and survival rate after surgical treatment were detailed. The cats included 11 domestic shorthair (DSH) and three Persian. Trauma was the most common cause of rupture in these patients and one 3-month old Persian cat had peritoneopericardial diaphragmatic hernia (PPDH). The most common clinical sign was dyspnea. Concurrent disorders included fractures of pelvic, long bones and ribs and mild pneumothorax. One of the five cats survived with concurrent disorders. In this study, the frequency of rupture sites was as follows: the right-side (11 cats), left-side (2 cats), and dorsal side of diaphragm in one case. The most frequently inserted organ into the chest was liver. In our study, the survival rate after surgical treatment was 71.00%. In conclusion, we suggested that stabilization of the patient was important before herniorrhaphy. Based on our study, the concurrent damages affected the survival rate.

Abstract Different living organisms are used as applicable bioindicators to determine heavy metal pollutions. Recent studies have shown that helminths parasites can be used as efficient environmental sentinels. This study aimed to evaluate Fasciola hepatica and Dicrocoelium dendriticum as bioaccumulators of lead (Pb), chromium (Cr), cadmium (Cd), copper (Cu). For this work. A total of 50 samples (F. hepatica, D. dendriticum, and livers from the infected and uninfected sheep, each of 10 samples) were collected from sheep slaughtered in Tabriz abattoir. One gram of each sample was incinerated and analyzed by Flame Atomic Absorption Spectrometry. The analysis of samples showed that Pb, Cr and Cu values in F. hepatica were higher than those in D. dendriticum, but only the differences of Pb and Cu were significant. The values of heavy metals in F. hepatica were significantly higher than those in the infected livers (except for Cd), while in D. dendriticum, Cr and Cd were only higher. Based on metal levels in livers, it was found that bioconcentration factors (BCFs) of Cr, Pb and Cu for F. hepatica were much more than one, and BCFs of these three metals between two flukes were statistically significant. This study indicated that F. hepatica had a higher bioindicator potential than D. dendriticum to evaluate environmental pollutants by some metals.

Abstract A one and a half years old male French bulldog weighing 9.50 kg was presented with the history of inappetence, lethargy, abdominal distension and exercise intolerance since last 2 days. The physical examination was done which revealed normal physiological parameters including temperature, mucus membrane color and capillary refill time except palpable precordial thrills, jugular distension on palpation, tachycardia and systolic murmurs on auscultation. Electrocardiography (ECG) was done which depicted ectopic foci with atrioventricular junctional tachycardia and right ventricular enlargement involving very small inverted P waves, deep S waves in leads I, II, III and augmented vector foot (aVF) and splintered QRS complexes. The dog was undergone chest radiography that revealed right atrial enlargement, increased sternal contact of heart on lateral view and a bulge at 9:00 o’clock to 11:00 o’clock depicted right atrial enlargement on dorso-ventral view. Lastly, echocardiography was done to arrive at a diagnosis confirming the Ebstein’s anomaly as a form of tricuspid valve dysplasia including apical displacement of tricuspid valve leaflets, division of right ventricle into atrialized and functional portions, increased displacement index, increased apex-mitral annulus to apex-tricuspid annulus ratio, severe right atrial dilatation and tricuspid regurgitation. The dog was medically treated with diuretics, angiotensin converting enzyme inhibitors and inotropes and the owner was advised to put the dog on low sodium diet for 2 weeks. The dog has resolved clinical signs of right sided heart affection; but, suddenly collapsed at home. The owner denied for the necropsy of dog.

Abstract The development of tooth is a complex process wherein there is a series of interactions between the ectoderm and ectomesenchyme. Hyperdontia (also called supernumerary teeth) is a developmental abnormality being additional to the normal series of teeth with unknown etiology and is rare in ruminants. In this report, a rare case with two abnormal teeth outside the alveolar region, observed in a healthy 3-month-old female lamb was presented. The congenital defect consisted of the presence of 2 teeth positioned bilaterally in the caudal part of the mandible ramus and under the prominent pendulous skin; so, with retraction of prominent pendulous skin, these ectopic teeth were seen outside of the skin. The anomaly was found during a routine physical examination and no other congenital abnormalities were found in the lamb. The etiology of supernumerary teeth is not completely understood. This odontogenic abnormality appears to be an extraordinarily rare condition. Genetic, toxic, infectious or environmental factors related to congenital abnormalities affect the tooth growth. Depending on the supernumerary teeth position, surgery may be required.