Association of the severity of colic in horses with oxidative stress biomarkers, acute-phase proteins, and certain trace elements.

Sixty-one horses were enrolled in this study and divided into 3 different groups according to their severity of colic (heart rate, oral mucous membrane color, and abdominal distention): a strangulating colic (SC) group (n=21), non-strangulating colic (NC) group (n=20), and control group (n=20) consisting of randomly selected normal horses without signs of colic. The serum concentrations of haptoglobin, tumor necrosis factor-α (TNFα), nitric oxide (NO), malondialdehyde (MDA), zinc, iron, and copper were evaluated in all horses. The average concentration of TNFα in the SC group was higher than that in the control group (P<0.001). The TNFα concentration was higher in the NC group compared with the control group (P<0.001). Furthermore, the average concentration of TNFα tended to be higher in the SC group compared with the NC group (P=0.052). The average concentration of haptoglobin in the SC group was higher than that in the control group (P<0.001). The average concentration of NO was higher in the SC group compared with the NC group. (P=0.016) The average concentration of MDA was higher in the SC group compared with the control group (P=0.042). Furthermore, the concentration of MDA was higher in the SC group compared with the NC group (P=0.048). TNFα in horses with signs of colic may be a reliable indicator of prognosis and the severity of clinical signs. The haptoglobin concentration may be a useful marker in cases where animals are referred to clinicians a few days after the onset of colic. The concentrations of MDA and NO should be interpreted with caution.

Serological survey of Toxoplasma gondii infection in the one-humped camels (Camelus dromedarius) in the south of Iran.

BACKGROUND: Toxoplasmosis, an important zoonotic disease, is caused by Toxoplasma gondii. Camels are one of several host species for T. gondii parasites and play an important role in the transmission of T. gondii to humans. OBJECTIVES: The present study aimed to describe the seroprevalence of T. gondii in dromedary camels (Camelus dromedarius) from three provinces (Fars, Bushehr and Hormozgan), southern Iran first for this host. METHODS: A total of 180 serum samples were analysed for the presence of anti-Toxoplasma IgG antibodies using the enzyme-linked immune-sorbent assay. RESULTS: Our results showed an overall seroprevalence of T. gondii in 15% of animals. Antibodies to T. gondii were found in sera of 27 of 180 dromedary camels from Fars, Bushehr and Hormozgan provinces, southern Iran. Age or the gender of the camel did not significantly affect the seroprevalence (p > 0.05). There was no significant association between herd-level seroprevalence of T. gondii infection and abortion history, province location residence, history of animal keeping and history of contact with other animals (p > 0.05). CONCLUSIONS: The results of this study showed the presence of T. gondii antibodies among camels in Southern Iran, which could be a public health concern. According to the prevalence of T. gondii infection in camel, the implementation of control measures to reduce infection in both definitive and intermediate hosts is needed.

Molecular and hematological investigation of Trypanosoma evansi infection in Iranian one-humped camels (Camelus dromedarius).

Trypanosoma species cause animal trypanosomiasis that infects many animals. Trypanosoma evansi is an organism that infects camels. There are many economic problems associated with this disease, including lower milk and meat yields and abortions. The purpose of the current survey was molecular study of the presence of Trypanosoma in dromedary camel blood in the south of Iran, and its effects on the hematologic, and some acute-phase protein changes. Blood samples were aseptically collected from the jugular vein of dromedary camels (n = 100; aged from 1 to 6 years) originating from Fars Province in EDTA-coated vacutainers. Genomic DNA from 100 µL of the whole blood was extracted and amplified using a PCR assay based on ITS1, 5.8S, and ITS2 ribosomal regions. Also, the PCR products obtained were sequenced. Moreover, the changes in hematological parameters and serum acute-phase proteins (serum amyloid A, alpha-1 acid glycoprotein, and haptoglobin) were measured. Among 100 tested blood, nine samples (9%, 95% CI: 4.2-16.4%) were found positive by the PCR assay. The phylogenetic tree and blast analysis showed four different genotypes closely related to the strains (accession numbers: JN896754 and JN896755) previously reported from dromedary camels in Yazd Province, center Iran. Based on hematological analysis, normocytic and normochromic anemia and lymphocytosis were detected in the PCR-positive cases compared with the negative group. Furthermore, alpha-1 acid glycoprotein was significantly increased in the positive cases. There was a substantial and positive relation between the number of lymphocytes, and the levels of alpha-1 acid glycoprotein and serum amyloid A in the blood (p = 0.045, r = 0.223 and p = 0.036, r = 0.234, respectively). A noticeable frequency of T. evansi infection was reported in dromedary camels in south Iran. This is the first report on the genetic diversity of T. evansi in this region. There was a significant association among Trypanosoma infection, lymphocytosis, and alpha-1 acid glycoprotein. Trypanosoma-positive camels had a significant decrease in hematocrit (HCT), hemoglobin (Hb), and red blood cell (RBC) values compared to the non-infected group. Further experimental studies are needed to elucidate the hematological and acute-phase protein alteration during a different phase of Trypanosoma spp. infection.

Theileria equi in the horses of Iran: Molecular detection, genetic diversity, and hematological findings.

In all equids worldwide, Theileria equi and Babesia caballi are believed to be two important erythrocytic protozoa that cause equine piroplasmosis. In addition, it was recently discovered that Theileria haneyi is another potential equine piroplasmosis (EP) agent. Ixodid ticks are the major vectors of these parasites. Equine piroplasmosis is of international importance and affects enormously the equine industry. In this study, for the first time, molecular prevalence and genetic diversity of piroplasma parasites (T. equi and B. caballi) in horses from Fars province (south of Iran) were determined. Also, hematological alterations of naturally infected horses were analyzed. PCR positive horses showed anemia, thrombocytopenia, leukocytosis with a left shift of neutrophilia, and monocytosis. PCR results revealed that, from 133 blood samples of horses, 40 samples were positive (30.07%). The occurrence of T. equi in this area (30.07%) was more than the national average prevalence of T. equi (24.11%), but B. caballi prevalence in study area (0%) was less than the average of previous studies in Iran (5.47%). Our findings revealed that the T. equi was widespread in Fars province of Iran. PCR products of 18S rDNA and EMA-1 genes of T. equi strains were sequenced successfully. All 18S rDNA sequences collected in this experiment revealed 100% similarity together. According to the phylogenetic tree constructed using the 18S rDNA gene, Iranian T. equi is clustered with strains from Cuba (KY111762, KY111761) and USA (CP001669, JX177672). So, this could be concluded that T. equi studied in this research, and those strains are initiated from a common T. equi ancestor at an unknown time ago. Also, the phylogenetic tree based on EMA-1 gene demonstrated a genetically diverse population of Iranian T. equi strains (10 different genotypes). As EMA-1 is one of the most immunogenic antigens in this parasite, such variability could be a concern about the efficacy of T. equi vaccines. Finally, more studies on equine piroplasmosis in the provinces of the southern region of Iran are recommended to create a better vision of disease in this region.

First report of ocular histoplasmosis in a horse from Iran: molecular, clinical and pathological findings.

Histoplasma capsulatum is a dimorphic fungus that is traditionally classified in three varieties: Hc var. capsulatum, Hc var. duboisii, and Hc var. farciminosum (HCF). Cytology, hematology, pathology, polymerase chain reaction (PCR), sequencing, and phylogenetic analyses were applied on samples collected from the blood and the eye of a horse with pustular lesions and ocular discharge. Physical examination and cytopathological tests showed H. capsulatum infection. Additionally, the results of two PCR tests confirmed H. capsulatum infection. The phylogenetic tree of the internal transcribed spacer sequence of Iranian H. capsulatum showed homology with the HCF variety. For the first time, H. capsulatum infection in the eye of a horse from Iran was detected and phylogenetically analyzed. This study revealed that H. capsulatum could establish infection in Iranian animals in addition to people, and indicated the role of soil enriched with bird dropping in the preparation of a favorable environment for H. capsulatum propagation. Further investigations are required to clarify the natural history and risk factors associated with histoplasmosis in Iran.

Seroprevalence Study on West Nile Virus (WNV) Infection, a Hidden Viral Disease in Fars Province, Southern Iran.

BACKGROUND: West Nile Virus, a mosquito-borne flavivirus, causes a variety of symptoms in human, from asymptomatic infection to neuroinvasive disease. Several studies have been conducted on the seroprevalence of WNV infection in different areas from Iran. This study was performed to find the presence of antiviral antibodies in human serum among some high risk population and awareness of health care staff about symptom of the WNV infection. METHODS: Study performed in five geographical districts based on high population of immigrant and domestic birds and prevalence of the antiviral antibodies in horses which was reported previously. Totally 150 human blood samples were collected during 2018. The samples collected from patients referred to the clinics. The ELISA method used to detect IgG and IgM antibody against WNV. Logistic regression models used to analyze the effect of sex, age, keeping birds and urban/rural residence on the risk of infection. The awareness of health care staff about symptom of infection surveyed. RESULTS: From all blood donors, 41 samples (27.33%) showed positive to IgG antibody. From which 56.10% were males and remaining females. None of the mentioned factors had a significant relationship. Health care staff had less attention to the infection. CONCLUSION: Although the prevalence of antibodies was relatively high, due to the similarity to other viral diseases, health care staff had less attention to the disease. The study showed that people in these areas have been exposed to the virus. Further research activities are recommended for control of this arbovirus.

The occurrence of hemotropic Mycoplasma ovis-like species in horses.

Hemotropic mycoplasmas (haemoplasmas) are wall-less bacteria, which may lead to anemia and, even, mortality in mammals. The present study was aimed to characterize the causative agent(s) of haemoplasma infection in blood samples taken from horses (n = 133) in south of Iran. Microscopic examination of blood smears and PCR assay were performed for the detection of hemotropic Mycoplasma and equine piroplasma (Babesia caballi and Theileria equi). For the purpose of molecular characterization, 16S rDNA and 18S rDNA markers were used for hemotropic Mycoplasma and piroplasma pathogens, respectively. The PCR-positive samples were sequenced for haemoplasma and further phylogenetic analysis was performed for the obtained haemoplasma sequences. Nine out of 133 (6.77 %, 95 % CI: 2.5-11.04 %) horses were positive for Mycoplasma sp. by PCR. Furthermore, three of these animals were co-infected with T. equi. Interestingly, the phylogenetic and molecular analysis of the haemoplasma sequences derived from the PCR amplicons in the equine positive cases showed 100 % identity with 16S rDNA in Mycoplasma ovis, an HM member mainly found in sheep and goats. In addition, the hematological analysis showed that PCR positive horses for M. ovis-like species had a lower hematocrit (Hct), hemoglobin (Hb) concentration, and red blood cell (RBC) count compared to the PCR negative ones (P ≤ 0.05). Mild anemia was also developed in the haemoplasma-positive horses. These findings represent the first molecular evidence of M. ovis-like species in horses. Further experimental studies are needed to examine the importance of this nonspecific host infection and evaluate its pathogenicity in equine and other species.

'Candidatus Bartonella dromedarii' in the dromedary camels of Iran: Molecular investigation, phylogenetic analysis, hematological findings, and acute-phase proteins quantitation.

The genus Bartonella is comprised of Gram-negative coccobacilli, aerobic, and facultative intracellular bacteria which are transmitted by hematophagous vectors (e.g., fleas, lice, sandflies, and ticks). Each species of Bartonella infects one or few related mammals as reservoir host(s). If a Bartonella spp. infects a nonspecific host like humans, it can lead to a more acute disease. Bartonella spp. has been detected more recently for the first time in camels in Israel by Rasis and colleagues. However, the epidemiological and public health importance of this new pathogen in camels is not clear. In this study, we aimed to detect the Bartonella spp. in the blood samples of Iranian camels, measure their prevalence, and determine their species. Also, the relationship between Bartonella spp. infection and different hematological factors and acute-phase proteins (Hp, a1AGP, SAA) was investigated. Finally, the sequences of three DNA regions, i.e.16S rDNA, rpoB, and ITS, were determined and phylogenetically analyzed. From the 106 examined blood samples of camels from Fars province (southern area of Iran), 18 samples were positive (17%). The findings also showed that Bartonella spp. positive camels had significantly lower Hb, MCH, and MCHC but higher RDW, SAA, and WBC (P < 0.05) compared to the control group. Our Bartonella strain was genetically similar to the 'Candidatus Bartonella dromedarii' but different from Bartonella bovis. Thus, more studies are required to investigate the phenotypic and genotypic characteristics of 'Candidatus Bartonella dromedarii'. Also, there is a need to evaluate precisely the risk factors, transmission routes, and zoonotic potential of this species.

Application of polymerase chain reaction on cerebrospinal fluid for diagnosis of cerebral coenurosis in small ruminants.

Sheep and goats serve as intermediate hosts for the canine tapeworm Taenia multiceps. The cysts produced by the intermediate stage of parasite are usually found in the cerebral hemispheres of small ruminants, and the resulting disease is commonly known as coenurosis. Coenurosis is clinically manifested in the form of various nervous symptoms, depending on the exact location of the cyst. The variety of neurological symptoms contributes to the complexity of clinical diagnosis and reinforces the need for a more specific and acceptable diagnostic approach. We demonstrated here, for the first time, that the T. multiceps DNA is present in the cerebrospinal fluid (CSF) of the infected sheep and goats. In addition, the molecular genetic marker of the mitochondrial DNA was applied phylogenetically to show that our isolates together with other T. multiceps strains comprised a monophyletic group that is a sister to Taenia krabbei. Pairwise comparison between the cox1 sequences of our study and other T. multiceps genotypes existing in the GenBank showed similarity ranging from 98 to 100%. Accordingly, the polymerase chain reaction (PCR) can be used for amplification of DNA of the parasite originated from the CSF and provides a valuable method for accurate identification of coenurosis cases.