Seroprevalence and associated risk factors of Toxoplasma gondii in sheep in Erzurum province, Eastern Anatolia region, Turkey.

BACKGROUND: Toxoplasmosis causes economic losses due to abortion, neonatal death and reproductive diseases in infected sheep. Erzurum is one of the most important cities in Turkey where sheep farming is done. Aims: The aim of this study was to evaluate seroprevalence of Toxoplasma gondii and related risk factors in sheep brought to the slaughterhouse in Erzurum province of Eastern Anatolia region, Turkey. METHODS: Nine-hundred and sixty sheep brought to slaughterhouse from Erzurum center and districts were used in this study. The data on age, breed, abortion history of sheep and whether or not they had contact with cats were recorded. The presence of T. gondii antibodies was determined by enzyme-linked immunosorbent assay (ELISA) kit in blood samples taken from sheep just before slaughter. RESULTS: According to the study results, 44 (4.58%) of 960 sheep were found to be seropositive. Seroprevalence was found to be highest in the ≥2<3 age group with 5.29% (P>0.05), and it was more common in Akkaraman breed compared to Morkaraman breed (P<0.01). It was determined that 43 (97.73%) of the 44 seropositive sheep had contact with cats (P<0.01) and 12 of them (27.27%) had abort history. CONCLUSION: The study results identify the presence of T. gondii in the sheep from Erzurum province of Eastern Anatolia region, Turkey.

Molecular evidence for trans-stadial transmission of Anaplasma platys by Rhipicephalus sanguineus sensu lato under field conditions.

This study investigated the transmission of Anaplasma platys by Rhipicephalus sanguineus sensu lato (Ixodida: Ixodidae). Engorged nymphs (n = 404) removed from A. platys-infected dogs were incubated at 28 °C until moulting. Unfed adults were obtained and divided into 48 pools. Unfed ticks collected from the grounds of the dog shelter, comprising 1800 larvae (n = 18 pools), 3100 nymphs (n = 62 pools) and 85 adults (n = 10 pools, including three male and seven female pools) were sorted into 90 pools. All pools were screened by polymerase chain reaction for the 16S rRNA gene of A. platys. Of 48 pools of unfed adults obtained from engorged nymphs, 12 were positive for A. platys; the infection rate maximum likelihood estimate (MLE) was 3.36 [95% confidence interval (CI) 1.84-5.68]. Anaplasma platys was detected in five of 23 male pools (MLE 2.82, 95% CI 1.06-6.20) and seven of 25 female pools (MLE 3.83, 95% CI 1.72-7.57). Of seven pools of unfed adult females collected from the shelter grounds, one was positive for A. platys (MLE 1.74, 95% CI 0.11-8.22). Among 62 unfed nymph pools, eight were infected with A. platys (MLE 0.27, 95% CI 0.13-0.52). No A. platys DNA was detected in the larva pools. The present results reveal molecular evidence for the trans-stadial transmission of A. platys by R. sanguineus s.l.

The effect of sulforaphane on oxidative stress and inflammation in rats with toxic hepatitis induced by acetaminophene.

OBJECTIVE: The aim of the present study was to reveal the possible effect of sulforaphane on oxidative stress and inflammation in rats liver with toxic hepatitis induced by acetaminophene. BACKGROUND: Sulforaphane is a compound with high antioxidant properties. Acetaminophen, which is a para-aminophenol derivative, can lead to fatal hepatic necrosis with direct hepatotoxic effects at high doses. METHODS: Thirty six male Sprague-Dawley rats were randomly divided into four groups. Control group (n = 9) was fed with standard rat chow and water for 3 days. Group APAP (n = 9) received a single dose acetaminophen 1 g/kg by oral gavage in addition to standard chow and water. Group SFN (n = 9) received sulforaphane 500 µg/kg by oral gavage in addition to standard chow and water for 3 days. Group APAP+SFN (n = 9) received sulforaphane 500 µg/kg and a single dose acetaminophen 1 g/kg by oral gavage in addition to standard chow and water. Acetaminophen was administered three hours after SFN administration. RESULTS: Neopterin, MDA, AST, ALT and CRP levels of group APAP were significantly increased compared to control group. GSH level of group APAP was significantly lower than in the control group. CONCLUSION: Sulforaphane is a protective agent against acetaminophen-induced liver damage and it can be added in the treatment protocol (Tab. 1, Fig. 5, Ref. 51).

Molecular survey of haemoplasmas in shelter dogs and associations with Rhipicephalus sanguineus sensu lato.

This study aimed to assess the occurrence of canine haemoplasma infection in domestic dogs and its possible trans-stadial transmission by Rhipicephalus sanguineus sensu lato (Ixodida: Ixodidae) in shelter dogs in Diyarbakir Province in southeast Turkey. Blood samples (n = 282) collected from domestic dogs were analysed by polymerase chain reaction (PCR) for the presence of canine haemoplasma. Fully engorged nymphs (n = 204) were removed from dogs that were positive for canine haemoplasma by PCR and maintained in an incubator at 28 °C for moulting. Unfed ticks (n = 2185) comprising 2100 nymphs and 85 adults collected from the grounds of the same shelter were also screened. Of 282 dogs, 108 [38.3%, 95% confidence interval (CI) 32.6-44.2] were PCR-positive for canine haemoplasmas. Mycoplasma haemocanis (Mhc) infection (26.2%, 95% CI 21.2-31.8) was observed in a significantly higher number of dogs than was Candidatus Mycoplasma haematoparvum (CMhp) infection (6.7%, 95% CI 4.1-10.3). Co-infections were seen in 15 (5.3%, 95% CI 3.0-8.6) dogs. None of the tick specimens examined were found to be positive for haemoplasma. Partial sequences of the 16S ribosomal RNA (rRNA) gene shared 99-100% identity with the corresponding published sequences for Mhc and CMhp. The present results revealed no trans-stadial transmission of canine haemoplasma species by R. sanguineus s.l. in field conditions.

Transstadial Transmission of Hepatozoon canis by Rhipicephalus sanguineus (Acari: Ixodidae) in Field Conditions.

This study investigated possible transovarial and transstadial transmission of Hepatozoon canis by Rhipicephalus sanguineus (Latreille) ticks collected from naturally infected dogs in a municipal dog shelter and the grounds of the shelter. Four hundred sixty-five engorged nymphs were collected from 16 stray dogs that were found to be infected with H. canis by blood smear and PCR analyses and maintained in an incubator at 28 °C for moulting. Four hundred eighteen nymphs moulted to adults 14-16 d post collection. Unfed ticks from the shelter grounds comprised 1,500 larvae, 2,100 nymphs, and 85 adults; were sorted according to origin, developmental stage, and sex into 117 pools; and screened by 18S rRNA PCR for Hepatozoon infection. Of 60 adult tick pools examined, 51 were infected with H. canis. The overall maximum likelihood estimate (MLE) of infection rate was calculated as 21.0% (CI 15.80-28.21). Hepatozoon canis was detected in 31 out of 33 female pools (MLE 26.96%, CI 17.64-44.33) and 20 out of 27 male pools (MLE 14.82%, CI 20.15-46.41). Among 42 unfed nymph pools collected from the shelter, 26 were infected with H. canis, and MLE of infection was calculated as 1.9% (CI 1.25-2.77). No H. canis DNA was detected in any of the gDNA pools consisting of larva specimens. Partial sequences of the 18S rRNA gene shared 99-100% similarity with the corresponding H. canis isolates. Our results revealed the transstadial transmission of H. canis by R. sanguineus, both from larva to nymph and from nymph to adult, in field conditions. However, there were no evidence of transovarial transmission.

Molecular and Parasitological Survey of Ovine Piroplasmosis, Including the First Report of Theileria annulata (Apicomplexa: Theileridae) in Sheep and Goats from Turkey.

Blood and tick samples were collected from 333 apparently healthy sheep and 257 goats as well as 10 sheep exhibiting clinical signs of babesiosis in Adana, Gaziantep, and Adiyaman Provinces in southern Turkey. Fully engorged female ticks were selected and maintained in an incubator until they oviposited. The tick carcasses and their egg masses were examined. Piroplasms compatible with Babesia spp. and Theileria spp. were observed in both symptomatic and asymptomatic small ruminants. Genomic DNA isolates from blood of ovine, tick samples, and egg masses were screened for piroplasms by utilizing 18S rRNA polymerase chain reaction and reverse line blotting (RLB) assays. Parasitemia ranged from 0.01% to 5.6% of erythrocytes in clinical cases. RLB showed positivity in 239 (40.5%) of the sampled apparently healthy sheep and goats and revealed the presence of three Theileria and one Babesia species. Theileria ovis was the most prevalent (35.4%), followed by Babesia ovis (5.4%), Theileria annulata (3.9%), and Theileria sp. MK (0.3%). Thirty-two small ruminants infected with T. ovis were also infected with B. ovis One animal infected with T. ovis was also infected with Theileria sp. MK. Ticks were identified as Rhipicephalus bursa, Rhipicephalus turanicus, Hyalomma excavatum, Haemaphysalis parva, and Hyalomma anatolicum Egg masses of two female R. bursa carcasses were infected with B. ovis This is the first report of theileriosis caused by T. annulata in sheep and goats in Turkey.

Long-term follow-up and quality of life after robot assisted sacrohysteropexy.

OBJECTIVE: The aim of this study is to investigate the effect of robot assisted laparoscopic sacrohysteropexy (RALS), with preservation of the uterus, in patients with pelvic organ prolapse on short and long term outcome. We report on (anatomical) status of the prolaps and the associated health related quality of life of women treated with RALS before and five years after surgery. STUDY DESIGN: A prospective cohort study in a teaching hospital in The Netherlands was performed. Quality of life was assessed pre-operative, post-operative and five years after RALS using the UDI/IIQ validated self-questionnaire designed for Dutch-speaking patients. Clinical and operative data were prospectively collected up to five years. Statistical analysis of categorical data was performed with the paired T-test. Descriptive statistics were computed with the use of standard methods for means, median and proportions. RESULTS: Hundred women with utero vaginal prolapse were treated with RALS with preservation of the uterus. The overall success rate of pelvic organ prolapse (POP) was 89.2%. After surgery the quality of life improved (P<0.05) Overall health status, based on a 0-100% visual analogue scale (VAS), improved from 72.6% pre-operative to 82.2% six weeks postoperative (P<0.05). Postoperative patients experienced less feelings of nervousness (P=0.01), shame (P<0.05) and frustration (P<0.05). The positive effects on these feelings remained present after five years. The learning curve shows a decrease in operating time with gained experience. CONCLUSION: RALS has proven to be a safe and effective treatment for uterine preserving surgery in cases of pelvic organ prolapse. The long term anatomical outcomes and quality of life after RALS compare favorably with laparoscopic and open hysteropexy.

Molecular and Parasitological Survey of Bovine Piroplasms in the Black Sea Region, Including the First Report of Babesiosis Associated with Babesia divergens in Turkey.

Clinical cases of babesiosis were evaluated, and the frequency of bovine Babesia and Theileria parasites was determined in cattle. Blood samples and thin blood smears were collected from 23 cattle exhibiting clinical signs of babesiosis. In addition, tick and blood samples were collected from 100 apparently healthy cattle cograzing from the same area. Egg masses obtained from fully engorged female ticks were included. DNA isolated from blood and tick samples was screened for Babesia and Theileria by reverse line blot assay. Piroplasms compatible with Babesia spp. were observed microscopically for symptomatic cattle as circular, oval, elongated, or pear-shaped bodies. Parasitemia ranged from 0.08 to 0.9% for Babesia bovis, 2.5 to 15.4% for Babesia bigemina, and 7.4% for Babesia divergens. Reverse line blot showed positivity in 13 (13%) of the sampled clinically normal cattle and revealed the presence of three Babesia species. Babesia bovis was the most prevalent (9/100, 9%), followed by Babesia occultans (3/100, 3%) and B. bigemina (1/100, 1%). One animal infected with B. bigemina was also infected with B. bovis. The single animal infected with B. divergens showed symptoms of babesiosis. Ticks were identified as Rhipicephalus annulatus, Rhipicephalus turanicus, and Ixodes ricinus. One female R. annulatus and its egg mass were infected with B. bigemina. Neither Theileria annulata nor Theileria buffeli/orientalis infections were observed in cattle or ticks. This is the first report of clinical babesiosis caused by B. divergens in cattle from Turkey.

Application of the Reverse Line Blot Assay for the Molecular Detection of Theileria and Babesia sp. in Sheep and Goat Blood Samples from Pakistan.

BACKGROUND: The present study was designed to detect the presence of tick-borne parasites (Theileria and Babesia spp.) in 196 blood samples collected from apparently healthy sheep and goats from two provinces, Punjab and Khyber Pukhtoon Khwa, in Pakistan. METHODS: Reverse line blot (RLB) assay was applied for the parasitic detection by the amplification of hypervariable V4 region of the 18S ribosomal RNA (rRNA) gene. A membrane with covalently linked generic and species specific oligonucleotide probes was used for the hybridization of amplified PCR products. RESULTS: Parasites were detected in 16% of the ruminant blood samples under study. Two Theileria species, T. lestoquardi and T. ovis, were identified in samples. 25, of the total 32, infected animals were from Khyber Pukhtoon Khwa. CONCLUSION: Sheep were more prone to tick borne haemoprotozans as 81% infected samples were sheep as compared to 19% goats (P > 0.001). Risk factor analysis revealed that male (P = 0.03), animals infested by ticks (P = 0.03) and herd composed of sheep only (P = 0.001) were more infected by blood parasites.

A study on the determination of risk factors associated with babesiosis and prevalence of Babesia sp., by PCR amplification, in small ruminants from Southern Punjab (Pakistan).

Babesiosis is a parasitic infection due to the multiplication of tick borne parasite, Babesia sp., in erythrocytes of host, which includes a wide variety of vertebrates including small ruminants causing decreased livestock output and hence economic losses. The objective of the present study was to establish a PCR based method for the detection of Babesia sp. in small ruminant population in Southern Punjab and to determine the risk factors involve in the spread of babesiosis. A total of 107 blood samples were collected from 40 sheep and 67 goats in seven districts of Southern Punjab from randomly selected herds. Data on the characteristics of the animals and the herd were collected through questionnaires. 36 blood samples (34% of total) produced the DNA fragment specific for 18S rRNA gene of Babesia sp., by PCR amplification, of which 20 were sheep and 16 were goats. Samples from all seven district contained Babesia positive samples and prevalence varied between 18 to 68%. It was observed that male animals (P = 0.009) and young animals under one year of age (P = 0.01) were more prone to the parasite. It was observed that herds consist of more than 15 animals (P = 0.007), composed of mixed species of small ruminants (P = 0.022), associated with dogs (P = 0.003) and dogs having ticks on their bodies (P = 0.011) were among the major risk factors for the spread of babesiosis in small ruminants.

Isolation of pathogenic aerobic bacteria from the blood of septicaemic neonatal calves and the susceptibility of isolates to various antibiotics.

An automated blood culture system (BACTEC 9240) was used for the isolation of aerobic bacteria from the blood of septicaemic neonatal calves. Blood samples were collected from 90 clinically septicaemic and 20 healthy neonatal calves and inoculated into blood culture bottles. There were 89 significant isolates from 90 positive blood cultures using the BACTEC system. Escherichia coli was the most common pathogen detected accounting for 56 (63%) out of 89 isolates. The other pathogens were beta-haemolytic streptococci (15.7%), Staphylococcus aureus (10.1%), Klebsiella sp. (5.6%) and Corynebacterium sp. (5.6%). All isolates showed a susceptibility rate of 100% to enrofloxacin, cefepim, cefoperazone/sulbactam, imipenem and meropenem while some of them were ranged from 75 to 91.7% susceptible to amoxicillin/clavulanic acid, ampicillin/sulbactam, gentamicin and cephalosporins.

Molecular identification of ovine Theileria species by a new PCR-RFLP method.

Theileria spp. infect wild and domestic ruminants in the tropical and subtropical regions of the world. Two species, T. lestoquardi and T. ovis, are suspected to cause ovine theileriosis in Iran. The epidemiological aspects of ovine theileriosis in Iran are poorly understood, and further investigations by sensitive and precise techniques are required. In this study, the use of a nested PCR for amplification of a fragment of the 18S ribosomal DNA from virtually all species of Theileria is described. For differentiation of various Theileria spp. a RFLP assay was developed as a diagnostic tool enabling direct, concurrent, highly specific and sensitive identification of Theileria spp. The sensitivity of the nested PCR for Theileria species was 10(-5)% parasitemia. Restriction fragment length polymorphism (RFLP) of the PCR products allowed differentiation between three different Theileria species (T. annulata, T. lestoquardi and T. ovis) and seems to be useful for differentiation of other species such as T. separata and Theileria spp. china. From 100 field blood samples obtained from sheep in East and South-East Iran, 56% were positive for Theileria spp. by nested-PCR compared with 21% by microscopic examination. Out of 56 positive samples, 12.5% (7/56) were positive for T. ovis and 87/5% (49/56) were positive for T. lestoquardi. This is the first report in which T. ovis has been detected in Iran using molecular identification techniques.

The perceived role of Islam in immigrant Muslim medical practice within the USA: an exploratory qualitative study.

BACKGROUND: Islam and Muslims are underrepresented in the medical literature and the influence of physician's cultural beliefs and religious values upon the clinical encounter has been understudied. OBJECTIVE: To elicit the perceived influence of Islam upon the practice patterns of immigrant Muslim physicians in the USA. DESIGN: Ten face-to-face, in-depth, semistructured interviews with Muslim physicians from various backgrounds and specialties trained outside the USA and practising within the the country. Data were analysed according to the conventions of qualitative research using a modified grounded-theory approach. RESULTS: There were a variety of views on the role of Islam in medical practice. Several themes emerged from our interviews: (1) a trend to view Islam as enhancing virtuous professional behaviour; (2) the perception of Islam as influencing the scope of medical practice through setting boundaries on career choices, defining acceptable medical procedures and shaping social interactions with physician peers; (3) a perceived need for Islamic religious experts within Islamic medical ethical deliberation. LIMITATIONS: This is a pilot study intended to yield themes and hypotheses for further investigation and is not meant to fully characterise Muslim physicians at large. CONCLUSIONS: Immigrant Muslim physicians practising within the USA perceive Islam to play a variable role within their clinical practice, from influencing interpersonal relations and character development to affecting specialty choice and procedures performed. Areas of ethical challenges identified include catering to populations with lifestyles at odds with Islamic teachings, end-of-life care and maintaining a faith identity within the culture of medicine. Further study of the interplay between Islam and Muslim medical practice and the manner and degree to which Islamic values and law inform ethical decision-making is needed.

Separation of adult bone marrow mononuclear cells using the automated closed separation system Sepax.

BACKGROUND: The Düsseldorf-based cardiologist Professor Strauer was the first to present a therapeutic concept for the repair of acute infarcted myocardium in 2001: the autologous intracoronary transplantation of unfractionated human bone marrow (BM) mononuclear cells (MNC). The Division of Cardiology, Pneumology and Angiology, University of Duesseldorf Medical School, Duesseldorf, Germany, was also able to show the regenerative potential of BM stem cell transplantation in patients with chronic heart disease (CHD) and peripheral arterial disease (PAD). In the mean time, several clinical trials have been set up worldwide, predominantly by using MNC isolated manually from BM aspirates via density-gradient centrifugation; 374 patients have been treated here with unselected BM MNC since 2001. Altogether 217 BM aspirates have been processed manually. In order to maintain the high standards required for cellular therapeutics, the Sepax cell-separation system was implemented into routine BM processing in 2006. The closed Sepax system provides a reproducible MNC isolation method, and 157 BM samples have been processed with the Sepax device. The results of manual MNC isolation were compared with the Sepax-mediated MNC isolation. METHODS: The manual Ficoll separation method was compared with the Sepax density gradient-based separation (DGBS) protocol using Ficoll with the kit CS-900 and the Sepax S-100 main processing unit from Biosafe. RESULTS: Nucleated cell and MNC recovery were significantly higher after Sepax processing (P<0.0001) whereas no significance was found for red blood cell depletion. DISCUSSION: The Sepax cell-separation system is a time-saving method providing clinical-grade MNC isolated automatically from human BM by Ficoll density centrifugation.

Genetic analysis of the M RNA segment of Crimean-Congo hemorrhagic fever virus strains in Turkey.

Crimean-Congo hemorrhagic fever (CCHF) virus is member of the genus Nairovirus of the family Bunyaviridae. All members of the family Bunyaviridae are enveloped viruses containing tripartite, negative polarity, single-stranded RNA. CCHF is characterized by high case mortality, occurring in Asia, Africa, the Middle East and Europe. During recent years, outbreaks have been reported in Turkey. However, little information is available on the genetic diversity of CCHF virus in Turkey. In this study, a total of 1227 adult ticks were collected from domestic ruminants (796 specimens from cattle, 399 specimens from goats and 32 specimens from sheep). The presence of the M segment of CCHF virus was determined in 4 of 40 (10%) Hyalomma marginatum marginatum pools, in 2 of 38 (7.89%) Rhipicephalus bursa pools, and in 1 of 7 (7%) Boophylus annulatus pools. Hyalomma anatolicum anatolicum pools gave negative RT-PCR result against CCHF virus. Serum samples from seven patients infected with CCHF were selected and subjected to RT-PCR to amplify partial M segment of CCHF virus. This report introduces the first data on partial nucleotide sequences of M RNA segments of CCHF virus strains circulating in Turkey, isolated from ticks.

Detection of Babesia ovis by PCR in Rhipicephalus bursa collected from naturally infested sheep and goats.

Polymerase chain reaction (PCR) was used to assess the presence and the frequency of Babesia ovis infection in the adult Rhipicephalus bursa and their hosts in Elazig province located in eastern Turkey. Tick and blood samples were collected from 32 sheep and 28 goats of four selected herds. A total of 226 R. bursa were randomly selected from the collected ticks and their salivary glands were dissected out in 0.85% saline under stereo microscope. DNA amplification method revealed that the frequency of B. ovis infections in the ticks and the small ruminants were 16.37% (37/226) and 6.66% (4/60), respectively. Three positive products, two of which were from the salivary glands of R. bursa and the other from sheep blood were purified from agarose gel and sequenced. The results showed that nucleotide sequences were identical to the previously reported nucleotide sequences of B. ovis. It is concluded that R. bursa might play an important role in the field as a natural vector of the parasite.

PCR-based detection of Theileria ovis in Rhipicephalus bursa adult ticks.

Tick-borne diseases in ruminants are common in tropical and subtropical regions and lead to meat and milk production losses. In this study, polymerase chain reaction (PCR) was used to assess the presence of Theileria ovis in Rhipicephalus bursa ticks. We have demonstrated that the PCR enabled detection of T. ovis in field isolates of R. bursa collected from naturally infested sheep and goats in eastern Turkey. The sampling was done in spring season (between May and June 2004). A total of 420 R. bursa were collected and randomly selected 192 number of them (97 female and 95 male) were dissected. Primers specific for 520 bp fragments small subunit ribosomal RNA (ssu rRNA) gene of T. ovis amplified products from 37 of the 192 (19.27%) samples. The parasite was detected in 17 (17.52%) female and in 20 (21.05%) male ticks. Two T. ovis amplicons from the tick samples were purified and sequenced. The resulting sequences were identical to the nucleotide sequence of the Turkish sheep strain of T. ovis. These results showed that R. bursa might play an important role in the field as a natural vector of T. ovis.