Seroprevalence and risk factors of brucellosis in Arabian horses.

BACKGROUND: Brucellosis, as a zoonotic disease, mainly occurs in horses by Brucella abortus, Brucella canis and Brucella suis. The disease in equines is often asymptomatic, but the clinical signs in horses are mostly characterized by bursitis, arthritis and tenosynovitis. OBJECTIVES: This study, thus, aimed to determine the seroprevalence of brucellosis and its associated risk factors in the Arabian horses of Khuzestan province, South-west Iran. METHODS: To that end, the blood samples randomly collected from 180 Arabian horses were analyzed for the presence of anti-Brucella antibodies by Rose Bengal plate test (RBPT), serum agglutination test (SAT), 2-mercaptoethanol test (2-ME) and a commercial i-ELISA kit. RESULTS: The ROC curve analysis showed that the best cut-off point for S/P values in i-ELISA turned out to be 26.25%. The results showed that the overall seroprevalence of brucellosis based on parallel interpretation of the test results was 12.22% (Positive/Tested = 22/180). The prevalence of acute and chronic brucellosis was 8.3 and 3.9%, respectively. The seroprevalence of brucellosis with RBPT and i-ELISA methods was 1.11% (2/180) and 7.22% (13/180), respectively. According to what SAT revealed, 9.44% (17/180) of sera had a titer of 40 or greater, and at 2-ME, 7.22% of samples (13 out of 180 samples) depicted a titer of 40. The results of i-ELISA, SAT and 2-ME were significantly different from those of RBPT (p < 0.01); however, there was no significant difference between i-ELISA, SAT and 2-ME in findings (p > 0.05). CONCLUSIONS: The results of this study recommend that i-ELISA be used for screening purposes of brucellosis in horses. The findings confirmed that Arabian horses are natural hosts for the Brucellae. It is, thus, necessary to adopt appropriate prevention and control programs by health authorities and horse owners so as to reduce the distribution and transmission of the infection in the regions where brucellosis is prevalent.

An exploratory study on the presence of Helicobacter heilmannii and Helicobacter billis in the feces of companion dogs.

BACKGROUND: Companion animals like dogs play an important role in the lives of many people and are often considered to be members of families, but definitely, any contact with them poses an inherent risk of transmitting zoonotic pathogens. One of these pathogens is the genus Helicobacter which is linked to many disorders in human and animal. OBJECTIVES: The aim of this study was to investigate the presence of some zoonotic species of genus Helicobacter in companion dogs. RESULTS: Through culturing in a special medium, nine samples (9%) were detected as infected (two pure and seven mixed culture). Based on multiplex-PCR, 13 samples (13%) were infected by Helicobacter spp. although none of them were infected by H. pylori. Species-specific PCR indicated that 38.5% or 5/13 of the samples were infected with H. heilmannii, while 15.45% or 2/13 of the samples were infected by H. billis. Multivariate logistic regression analysis showed that the age factor had a significant effect on Helicobacter spp. infection (odds ratio [OR] = 2.42, p = 0.01). CONCLUSION: This study revealed the negligible faecal transmission of H. pylori. Moreover, due to the detection of H. Heilmannii and H. billis in feces and their association with human gastric diseases, dog owners should be educated about the risks and transmission modes of zoonotic bacterial infections of dogs.

Comparison of Mycobacterium avium subsp. paratuberculosis infection in cattle, sheep and goats in the Khuzestan Province of Iran: Results of a preliminary survey.

BACKGROUND: Paratuberculosis or Johne's disease, the chronic infectious granulomatous enteritis of ruminants, is a worldwide infection, which is caused by Mycobacterium avium subsp. paratuberculosis (MAP). The most common symptoms of this disease in cattle are loss of milk production, weight loss and diarrhoea, whereas in sheep and goats, the symptoms are emaciation, anorexia and severe disability. OBJECTIVES: The aim of this study was to compare the seroprevalence of MAP in cattle, sheep and goats in the southwest of Iran. METHODS: Blood samples were randomly collected from 530 cattle, 568 sheep and 368 goats in southwest of Iran. Sera were tested by a commercial ELISA kit (ID vet; ID Screen® Paratuberculosis Indirect) for detection of antibodies of MAP. RESULTS: Overall apparent and true seroprevalence rate of MAP was 6.00% (95% CI: 4.90%-7.30%) and 13.25% (95% CI: 11.55%- 14.95%). Apparent and true seroprevalence of MAP, respectively, was 4.34% (95% CI: 3.88%-6.46%) and 9.19% (95% CI: 6.98%-11.98%) in cattle, 6.87% (95% CI: 5.05%-9.27%) and 15.37% (95% CI: 12.60%-16.60%) in sheep and 7.07% (95% CI: 4.82%-10.18%) and 15.86% (95% CI: 12.41%-20.01%) in goats, respectively. As a result, there was no significant relationship between animal species and MAP infection. Moreover, multivariate logistic regression showed that the infection rate is not associated with age, gender and geographical location in cattle, sheep and goats (P > 0.05). CONCLUSION: This study confirms that the seroprevalence of MAP is relatively considerable in the cattle, sheep and goats in the southwest of Iran, although in cattle, it is less than goats and sheep. Therefore, preventive and control measures should be considered by animal health authorities and meat and dairy processing units.

Crocin mitigated cognitive impairment and brain molecular alterations induced by different intensities of prenatal hypoxia in neonatal rats.

INTRODUCTION: Brain hypoxia has important role to the onset and progression of sporadic form of Alzheimer disease via expression of hypoxia-inducible factor-1 (HIF-1). Crocin by anti-amyloidogenic property inhibits ß-amyloid formation. However, the molecular mechanism associated with anti-amyloidogenic activity of crocin is unknown. So, the present study was designed to investigate the effect of crocin on cognitive behavior and expression of HIF-1α and ß-secretase (BACE1) genes in the brain of neonate rats following different intensities of hypoxia during pregnancy. MATERIAL AND METHODS: Pregnant female rats were divided into six groups including sham, control crocin treated (CC), hypoxia with three different intensities (H1-H3), and most intense of hypoxic group treated with crocin (H3C) (30 mg/kg; i.p) at P14. Hypoxia induced on the 20th day of pregnancy. Animals in sham and CC were put in hypoxia chamber at the same time of hypoxia group without any hypoxia induction. Morris water maze (MWM) and qRT-PCR were used to evaluate the cognitive behavior and mRNA levels of BACE1 and HIF-1α genes in the brain tissues. RESULTS: Animal under 7% O2  + 93% N2 condition for 3 hr showed the highest cognitive behavior impairment and upregulated HIF-1α and BACE1 mRNA in brains of offspring (p < .001). Crocin treatment improved memory impairment and attenuated the gene expression of HIF-1α and BACE1 in the brains of neonate rat. CONCLUSIONS: It was concluded that crocin has beneficial effects on the brain of neonate rats under gestational hypoxia by improvement of memory impairment and molecular alteration related to hypoxia.

Identification of Pseudomonas jessenii and Pseudomonas gessardii as the most proteolytic Pseudomonas isolates in Iranian raw milk and their impact on stability of sterilized milk during storage.

Identification of the most proteolytic Pseudomonas strains that can produce heat-resistant proteases and contribute to the Ultra High Temperature (UHT) milk destabilization is of great interest. In the present study, among the 146 Pseudomonas isolates that encoded the aprX gene, five isolates with the highest proteolytic activity were selected and identified based on 16S rRNA, rpoD and gyrB gene sequences data. The identification results were confirmed by phylogenetic analysis based on multilocus sequence analysis and identified the representative isolates as P. jessenii (two isolates) and P. gessardii (three isolates). Casein zymography demonstrated the ability of these species to produce heat-resistant enzymes, AprX, with molecular mass of about 48 kDa during storage at 7° C for 72 h. In sterilized milk samples, the residual activity of AprX caused a considerable enhancement in the degree of protein hydrolysis, non-protein nitrogen and non-casein nitrogen contents of the samples during a two-month storage. This enhancement was slightly higher in samples containing enzyme produced by P. jessenii compared to P. gessardii ones, resulting in earlier onset of sterilized milk destabilization. Hence, this study revealed that P. jessenii and P. gessardii can play a considerable role in deterioration of Iranian commercial long-life milk.

The Therapeutic Effects of Low-Frequency Electrical Stimulations Adjunct to Sodium Valproate on Seizure and Behaviors.

INTRODUCTION: Consuming antidepressant medications induce several problems leading to the need for alternative agents for emotional disturbances. Antidepressant medications increase the seizure risk; thus, alternative treatments, like Antiepileptic Drugs (AED), might be useful for patients with epilepsy comorbid with a psychiatric disorder. The present study evaluated the behavioral effects of sodium valproate, a none effective dose in seizure treatment [100 mg/kg; Intraperitoneal (IP)] along with the application of Low-Frequency Stimulations (LFS) during CA1 hippocampal kindling. METHODS: In total, 42 male rats were randomly divided into 6 groups, including control group with intact animals handled daily (I); sham group which was subjected to the surgical process, but received no real stimulation (II); saline-kindled Kindled group (S.kindled) which were stimulated daily with the following protocol: 3 strain of 50Hz monophasic pulses of 1ms duration applied 12 times a day with the threshold intensity at intervals of 10 minutes where saline was administrated 15 min before kindling stimulations (III); saline-kindled-LFS group (K4LFS) in which saline was injected 15 min before kindling stimulations and LFS was applied daily after the termination of kindling stimulation (IV); drug-kindle group (Drug100.kindled) that underwent rapid kindling procedure daily where sodium valproate (100 mg/kg) was administrated 15 min before kindling stimulations(V), and drug-kindled-LFS (Drug100.kindled.4LFS) group in which drug and LFS were administrated respectively before and after kindling stimulations (VI). The behavioral tests were assessed using elevated plus maze, open field, and forced swim tests. RESULTS: The combination of sodium valproate (100 mg/kg) and LFS significantly decreased cumulative seizure severity compared with the kindle group. Thus, it provided a strong seizure suppressing effect. Additionally, sodium valproate and LFS increased the percentage of Open Arms (OAs) entries and the OAs exploration; they also decreased jumping from elevated plus maze test and rearing in open field test. Furthermore, there was no significant change in the OAs entries and OAs exploration percentages, jumping from apparatus, and rearing in open field in Drug100. Kindled, K4LFS, and Drug100.kindled.LFS groups, compared with the sham group. There was no significant difference in the latency to first immobility and the duration of immobility in K4LFS groups compared with the S. kindled group. In the drug-kindled group, the latency to first immobility significantly increased, and the duration of immobility decreased, compared with the S. kindled group. Besides, the latency to first immobility significantly increased, and the duration of immobility decreased in drug-kindled-LFS, compared to S. kindled group; however, the latency to first immobility was not significantly changed, compared to drug-kindled groups. CONCLUSION: Sodium valproate and LFS can modulate the function of the brain regions involved in emotional processing in epilepsy, as well as anxiety- and depressive-like behaviors. Such a combination could also decrease emotional disturbances induced by the kindling process.

Evaluation of the pre-treatment effect of Centella asiatica medicinal plants on long-term potentiation (LTP) in rat model of Alzheimer's disease.

The present study was aimed to investigate the pre-treatment effect of Centella asiatica (CeA) extract on long-term potentiation (LTP) in a rat model of Alzheimer's disease (AD). A total of 32 male Wistar rats weighing 380 ± 30 g were randomly divided into four groups (n = 8). Group 1 (C: Control): the control group. Group 2 (L: Lesion): The nucleus basalis of Meynert (NBM) of rats' brain was bilaterally destroyed by injection of Ibotenic acid. Group 3 (CeA): Animals in this group received the CeA leaf extract for only a period of six weeks. Group 4 (CeA + L): The NBM of rats was destroyed by Ibotenic acid after six weeks of a diet containing the CeA leaf extract. In all groups, LTP was recorded using the electrophysiological technique and fEPSP after high frequency stimulation (HFS). The results showed that the slope and amplitude of PS as well as the sub-curve level significantly increased in the CeA + L group compared with the L and CeA groups. The CeA extract improved and strengthened the slope, amplitude and sub-curve surface of cumulative waves in animals with NBM lesion. The results showed that administration CeA extract for six weeks before induction of NBM lesion and induction of Alzheimer could enhance memory. In other words, the CeA extract had a preventive or protective role. The present study showed that CeA had a protective role for neurons among rats with NBM lesion.

Crocin Treatment after Maternal Hypoxia Attenuates Spatial Memory Impairment and Expression of BACE1 and HIF-1α in Rat Offspring Brain.

INTRODUCTION: Hypoxia via expression of Hypoxia-Inducible Factor-1 (HIF-1) is an important and effective factor in the onset and progression of memory disorders, such as Alzheimer Disease (AD). The activity of ß-secretase (BACE1) is increased in hypoxia conditions. BACE1 triggers a cascade of pathological events resulting in AD. Crocin acts as a memoryimproving agent but its molecular mechanism is not well-known. Therefore, in this study, the effect of crocin on spatial memory, HIF-1α, and BACE1 gene expression was investigated in rat offspring under maternal hypoxia. METHODS: Female pregnant rats on the 20th day of pregnancy were divided into 4 groups, including sham, crocin-treated, hypoxia, and hypoxia group treated with crocin. In the hypoxia groups, pregnant rats were exposed to 7% oxygen and 93% nitrogen intensity for 3 h. In the crocin-treated group, crocin (30 mg/kg) was injected at P14-28 (i.p). At the end, Morris water maze was used to assess spatial memory and real-time polymerase chain reaction was performed to measure the expression of BACE1 and HIF-1α genes in the brain of offspring. RESULTS: Maternal hypoxia impaired memory compared with the sham group. However, crocin treatment improved cognitive behavior. HIF-1α and BACE1 expressions were upregulated in the brain of offspring in the hypoxia group. Crocin treatment could attenuate the expression of both genes. CONCLUSION: According to our results, down-regulation of HIF-1α and BACE1 gene expressions in the brain of rat offspring after crocin treatment can be suggested as a molecular mechanism for crocin to improve spatial memory.

The role of sheep in the epidemiology of Bovine alphaherpesvirus 1 (BoHV-1).

Bovine alphaherpesvirus 1 (BoHV-1) as a cusative agent for some diseases in cattle infects sheep and goat; and it is believed that these animals may be reservoir host for this virus. Thus, BoHV-1 infection in sheep and goat should be considerd when there is a program for control and eradication of this virus in cattle. Therefore, the aim of this study was to determine the seroprevalence of BoHV-1 in sheep, relationship between host and environmental factors with infection, and the role of sheep in the epidemiology of the BoHV-1. Blood samples were randomly collected from 310 healthy sheep in 6 cities of Khuzestan province (Southwest of Iran) including Ahvaz, Hendijan, Shushtar, Dezful, Masjed Soleyman and Behbahan. Sera were analyzed by virus neutralisation (VN) test for detection antibodies to BoHV-1. According to VN test, apparent and true seroprevalence seroprevalence of BoHV-1 infection was 28.4 % (95%CI: 23.4-33.4%) and 28.4 % (95%CI: 23.3-33.4%), respectively. Logistic regression revealed that the odds of infection between the age was 1.06 (95%CI: 0.9-1.25) (P > 0.05), implying that the odds of infection increased 6 % with rising one year of age. Besides, the relative frequency of infection in males was more than females', and the odds of infection in male sheep was identified to be 1.13 (95%CI: 0.47-2.71) (P > 0.05), compared to that in females. Moreover, in comparison to sheep with no history of abortion, the odds of infection in sheep with a history of abortion was 1.28 (95%CI: 0.57-2.87) (P > 0.05). The seroprevalence in Shushtar, Masjed Soleyman, Dezful, Ahvaz, Hendijan, and Behbahan were found to be 48.3, 46.7, 31.7, 20, 16.7, and 12 percent, respectively and 13.1 of fluctuation in infection can be justified by different geographical locations investigated in this study (P < 0.001). Considering the significant seroprevalence of BoHV-1, present study confirmed the role of sheep in the epidemiology of this virus and control of BoHV-1 in sheep should be considered by animal health authorities in areas where BoHV-1 is prevalent.

Morphometric development of sheep (Ovis aries) lymph nodes in fetal period.

Immune system plays crucial role in body and lymph nodes are essential parts of this system for combating pathogens. However, no study has ever been conducted on morphometric development of sheep lymph nodes in fetal period. Thus, this study attempted to examine the morphometric characteristics of a number of important lymph nodes of some lymphocenters of sheep during fetal period. To this end, 60 pieces of sheep fetuses collected from Ahvaz slaughterhouse were fixated in 10% formalin and then divided into four categories based on crown-rump length (CRL) following gender and weight determinations. Mandibular, caudal superficial cervical (prescapular), caudal mediastinum, jejunal mesenteric and popliteal lymph nodes were evaluated in five lymphocenters of head, neck, thoracic cavity, abdominal viscera and pelvic limbs, respectively. In each sample, nodes formation was visually checked and in cases of nodes formation, they were measured in terms of weight, length, width and thickness and collected data were statistically analyzed. The longest and shortest fetal CRLs were found to be 48.50 cm and 3.50 cm, respectively. Gender had no effect on study parameters in 32 male and 28 female fetuses. Study of sheep fetuses' lymph nodes revealed no macroscopic lymph node development by day 45, while all nodes were observable after the day 59. The shortest lymph node was mandibular node and the longest one was caudal mediastinum. Based on the results, it seemed that although the size of lymph nodes grows by age, this increase is not the same for all nodes and groups.

Association Study of rs1333040 and rs1004638 Polymorphisms in the 9p21 Locus with Coronary Artery Disease in Southwest of Iran.

BACKGROUND: Coronary artery disease (CAD) is a multifactorial and heterogenic disease. Recently, genome-wide association studies have reported that rs1333040 (C/T) and rs1004638 (A/T) single nucleotide polymorphisms (SNPs) in the 9p21 locus have very strong association with CAD. This study aimed to examine these associations in Southwest of Iran. METHODS: Blood samples were collected from 200 CAD patients and 110 healthy individuals with no CAD. The association of two SNPs with CAD was evaluated by PCR and restriction fragment length polymorphism. RESULTS: Chi-square test showed no association between rs1333040 SNP and CAD (X2: 4.66, df: 2, P=0.09). Also, there was no association between rs1004638 SNP and CAD (X2: 0.27, df: 2, P=0.88). CONCLUSION: No association was observed between rs1333040 and rs1004638 SNPs in the 9P21 region and CAD in Southwest of Iran.

Seroprevalence of Bluetongue in sheep in Kohgiluyeh and Boyer-Ahmad province, Iran.

Bluetongue (BT) is a viral disease of ruminants transmitted by Culicoides biting midges and has the ability to spread rapidly over large distances. The disease occurs almost worldwide between latitudes approximately 35˚ S and 50˚ N. Among the numerous diseases of ruminants, BT has gained considerable importance in recent years as one of the best examples of the effects of climate change on disease spread. Sheep are major livestock species in Iran, but studies of BT have not gained the priority compared to other diseases. Thus, the objective of this study was to describe the distribution and seroprevalence of bluetongue virus (BTV) infections in sheep in Kohgiluyeh and Boyer-Ahmad province of Iran, and to identify factors associated with the exposure of these sheep to BTV infection. Sera from 262 apparently healthy sheep were collected during the year 2011. The collected sera of the animals were screened with competitive enzyme like immunosorbent assay (c-ELISA). Two hundred and three (77.48%) out of 262 sera tested were positive to BTV antibodies. Statistically significant differences were found in the seroprevalence BT, between sex and age of sheep (p < 0.001). No statistically significant differences were observed in BTV seroprevalence among different seasons, nor among recently aborted and normally delivered.

Investigation of Possible Correlation between Giardia duodenalis Genotypes and Clinical Symptoms in Southwest of Iran.

BACKGROUND: Giardia duodenalis is one of the most important human enteric parasites throughout the world. Clinical symptoms of this parasite vary from asymptomatic infection to chronic diarrhea. Still it is not clear, whether different types of pathogenesis are due to different strains of organism or to variable host factors. The purpose of this study was to investigate possible correlation of clinical symptoms with assemblages among symptomatic and asymptomatic cases collected from southwest of Iran. METHODS: Fecal samples were collected from 100 symptomatic and asymptomatic cases, which were positive for G. duodenalis. The samples were subjected to semi-nested PCR and RFLP for gdh gene. RESULTS: Among symptomatic patients, 54% had mixed genotypes AII and BIII, 28% and 18% of samples indicated assemblages BIII and AII, respectively. In contrast, among asymptomatic cases, 64%, 26% and 10%samples had mixed genotypes, BIII and AII assemblages, respectively. Statistical analysis using Chi- Square test showed that there was no significant correlation between assemblage and clinical symptoms in current study. CONCLUSION: High prevalence of mixed infection in both groups may affect this conclusion, therefore further study in more details are necessary to clarify these finding. Additionally, it is important to carry out investigations regarding human host factors as well.