Histopatološka i molekularna dijagnoza sarkocistoze miokarda u ovaca i imunohistološka korelacija s parazitskim lezijama koje sadrže MMP-2 i MMP-9

This study was aimed at the histopathological and immunohistochemical level investigate cardiac tissue lesions caused by Sarcocystis spp. in sheep, and thereby, to collect detailed information on the presence of these parasites and the pathogenesis of sarcocystosis in ovine animals. PCR, a molecular diagnostic method, was used for the identification of the parasite species. Microscopic examination revealed that out of the 45 ovine cardiac tissue samples examined 23 contained Sarcocystis spp. cysts. The PCR analysis results showed that out of the 23 cardiac tissue specimens infected with sarcocysts, 7 were determined to be infected with Sarcocystis gigantea and 16 with Sarcocystis tenella. Immunohistochemical examination demonstrated that, emiquantitatively, MMP - 9 staining was stronger than MMP - 2 staining in the periphery of the parasite cysts.Cilj je ovog rada bio histopatološko i imunohistokemijsko istraživanje lezija srčanog tkiva uzrokovanih parazitom Sarcocystis spp. u ovaca te prikupljanje detaljnih informacija o prisutnosti ovih parazita i patogenezi sarkocistoze u ovaca. Molekularna dijagnostička metoda PCR upotrijebljena je za identifikaciju parazitskih vrsta. Mikroskopska analiza pokazala je da su od 45 uzoraka srčanog tkiva ovaca 23 sadržavala ciste parazita Sarcocystis spp. Na temelju rezultata PCR analize od 23 uzorka srčanog tkiva invadirana sarkocistama 7 uzoraka bilo je invadirano vrstom Sarcocystis gigantea, a 16 uzoraka vrstom Sarcocystis tenella. Imunohistokemijska pretraga pokazala je da je, semikvantitativno, MMP-9 obojenje bilo jače od obojenja MMP-2 na periferiji cista parazita

Could antiparasitic cure with clinoptilolite be achieved in dogs naturally infected with giardia duodenalis?

Bu çalışmanın amacı doğal zeolit bileşeni olan klinoptilolitin köpeklerde doğal olarak şekillenen giardiazise karşı antiparaziter etkinliğinin belirlenmesidir. Aydın bölgesinde bulunan doğal olarak Giardia duodenalis ile enfekte olan sahipli ve ishalli her iki cinsiyetten 6-44 aylık yaştaki 7 enfekte köpek çalışmada kullanıldı. İki ayrı gruba ayrılan çalışma materyali olarak I. gruptakilere 10 gün oral yolla 2 g/kg dozda klinoptilolit uygulanırken, II. grupta bulunan köpeklerin kontrol grubu olarak kalmaları sağlandı. Köpeklerde klinoptilolitin antiparazitik etkinliği 0. 3. 7. ve 10. günlerde rektal yolla alınan dışkı örneklerinin mikroskobik incelenmesiyle değerlendirildi. Gram dışkı başına kist miktarındaki azalma göz önünde bulundurulduğunda anti-giardial amaçla kullanılan klinoptilolitin 10. günde giardiazise karşı kist atılımında %99 etkinlik sağladığı belirlendi. Sağaltım grubunda sırası ile 0.,3., 7. ve 10. günlerde kist sayılarına ait geometrik ortalamalar 200552.4, 37.8, 113.3, 10.4 şeklinde belirlendi. kontrol grubunda ise 0. gün ile (195099,4) 10. günlerdeki (174436,1) kist atılımının geometrik ortalamalarındaki fazla değişimin göstermediği saptandı. Netice itibarı ile giardiazisli köpeklerde klinoptilolitin kist atılımını etkin şekilde azalttığı ve önerilen dozda herhangi bir yan etkiye neden olmaksızın kullanılabileceği söylenebilir.The aim of this study was to determine the antiparasitic efficacy of the natural zeolite component, clinoptilolite, against naturally occurring giardiasis in dogs. Seven infected dogs, aged 6-44 months of age, both sexually owned and diarrhea infected with Giardia duodenalis in the Aydın region were used in the study. Group I was administered clinoptilolite orally for 10 days at a dose of 2 g / kg. Group II dogs were provide to stay in the control group. The antiparasitic efficacy of clinoptilolite in dogs was evaluated by microscopic examination of stool samples collected at the beginning 0, 3rd, 7th and 10th day by rectal route. Considering the reduction in cyst excretion per gram feces, clinoptilolite used for anti-giardial purposes provided 99% efficacy in cyst excretion at 10th day. In the treatment group, the mean geometric averages of cyst counts were determined as 200552.4, 37.8, 113.3, 10.4, respectively, on days 0, 3, 7 and 10. On the other hand, in the control group, no significant change was observed in the geometric mean of cyst ruble excretion on day 0 (195099,4) 10 days (174436,1). It can be said that clinoptilolite effectively decreases cyst excretion in dogs with giardiasis and can be used in the recommended dose without causing any side effects

Cryptosporidium spp. in Dogs - Prevalence and Genotype Distribution

Background: Cryptosporidium spp. is a zoonotic protozoan parasite that affects the gastrointestinal tract of humans and animals. The disease can cause acute and chronic diarrhoea and even death in both humans and animals. In this study, it was aimed to determine the prevalence and genotype distribution of Cryptosporidiosis in shelter dogs in Diyarbakır province located in the Southeastern Anatolia Region of Turkey. Materials, Methods & Results: The animal material of the study consisted of 100 dogs of different breeds and sexes. Faecal samples were collected from the rectum with disposable latex gloves and placed in individual sample containers.  All of the samples were examined for Cryptosporidium  spp. by Kinyoun Acid Fast and Nested PCR methods. In the Kinyoun Acid Fast staining method, firstly, smear preparations were prepared from fresh faecal samples, fixed in pure methanol for 1 min and allowed to dry. The slides were kept in Kinyoun Carbol-Fuxin for 5 min, dipped in 50% ethyl alcohol, shaken, washed in tap water, kept in 1% sulphuric acid for 2 min and washed in tap water.  The slides were kept in methylene blue for 1 min, washed in tap water and allowed to dry. After drying, immersion oil was dripped and examined under a microscope at 100 magnification. DNA extraction was performed from all samples using GeneMATRIX Stool DNA Purification Kit according to the manufacturer's protocol. After Nested PCR analysis was performed. In the PCR step, primers 5'-TTCTAGAGCTAATACATGCG-3' and 5'- CCCATTTCCTTCCTTCGAAACAGGA-3' were used to amplify the 1325 bp gene region. In the nested PCR step, primers 5'- GGAAGGGTTGTATTTATTTATTAGATAAAG-3' and 5'-AAGGAGTAAGGAACAACCTCCA-3' were used to amplify the 826-864 bp gene region. As a result of both methods, a prevalence of 3% was determined. The infection rate was higher in males (3.57%) than females (2.27%) and in younger than 1 year (5.56%) than in older than 1 year (1.56%). The DNA sequences obtained from the sequence analysis of 3 positive PCR samples were analysed in BioEdit software. A phylogenetic tree was constructed with the data set created by using the 18s rRNA gene sequences obtained from the NCBI genbank database and the DNA sequences obtained as a result of the study, and it was shown which Cryptosporidium species the study samples were related to. Today, many Cryptosporidium species have been identified and most of these species have host adaptation. Although C. canis is the most common species in dogs, C. muris, C. meleagridis, and C. parvum have also been detected. Among these species, C. parvum is recognized as a zoonotic species infecting a wide range of mammals. In this study, DNA sequencing of nested PCR positive samples revealed that 3 samples were zoonotic C. parvum. Discussion: This suggests that dogs may be a reservoir for zoonotic transmission of Cryptosporidium.  Consequently, it is recommended that people should be informed about the potential for transmission of this protozoan to humans and animals and that control programmes should be implemented, including the prevention of free entry of stray dogs into public places and homes. Keywords: Cryptosporidium  parvum, molecular analysis, canine, Diyarbakır, Turkey

Application of oral clinoptilolite intended for prevention of giardiasis in goat kids

Bu çalışma doğal zeolit olan klinoptilolitin oğlaklarda doğal yolla oluşan giardiazise karşı terapötik etkinliğinin belirlenmesi amacıyla gerçekleştirildi. Aydın ilinde yer alan bir keçi işletmesinde ishal anamnezi bulunan G. duodenalis ile doğal enfekte her iki cinsiyetten ve 19-41 günlük yaştaki 16 oğlak çalışma kapsamına alındı. Oğlaklar her grupta n=8 olacak şekilde randomize olarak 2 ayrı gruba ayrılarak I. gruba 1 g/kg dozda 10 gün oral yolla klinoptilolit uygulanırken, II. gruptaki olgulara hiçbir ilaç uygulaması yapılmayarak kontrol grubu olarak bırakıldı. Oğlaklarda klinoptilolitin terapötik etkinliği 0. ve 10. günlerde rektumdan direkt olarak toplanan dışkı numunelerinin mikroskobik muayenesiyle değerlendirildi. Gram dışkıda kist atılımındaki azalma değerlendirildiğinde klinoptilolit giardiazise karşı 10. günde kist atılımında %95 etkinlik sağladı. Buna karşın kontrol grubunda 0. gün ile (165832) 10. günlerde (162096) kist atılımının geometrik ortalamalarındaki azalmanın yeterli olmadığı görüldü. Sonuç olarak giardiazisli oğlaklarda oral yolla 10 gün uygulanan klinoptilolitin kist atılımını etkin şekilde azaltılmasına yardımcı olduğu söylenebilir.The present study was established in an attempt to interpret treapeutic efficacy of clinoptilolite against naturally occurring giardiasis in goat kids. A total of sixteen goat kids naturally infected with G. duodenalis at the age of 19 to 41 days, of both sexes were enrolled into the study from a goat farm located in Aydin province. Goat kids were randomly assigned into two groups (n=8), whereas group I received oral clinoptilolite at a dosage of 1 g/kg for 10 days, goat kids in group II were left as control without receiving any drug. Therapeutic efficacy of clinoptilolite was determined by microscopical interpretation of faecal samples collected from the rectum directly on days 0 and 10. Interpretation of the percentage reduction in cyst excretion revealed clinoptilolite efficacy as 95% on day 10. Indeed geometric means of the number of excreted cysts were not reduced sufficiently in control group between day 0 (165832) and 10 (162096). In conclusion, it might be suggested that oral administration of clinoptilolite for 10 days may have helped significant reduction the excretion of cysts in goat kids with giardiasis

Molecular Survey of Toxoplasma gondii Infection in Aborted Fetuses of Sheep in the Iğdır Province of Türkiye

Toxoplasma gondii, an obligatory intracellular protozoan parasite, can infect a wide range of warm-blooded animals, including livestock species. T. gondii is a zoonotic protozoan parasite that affects both humans and other warm-blooded animals. The aim of this study was to detect T. gondii by using PCR in the brain tissues of 60 aborted sheep fetuses from the Iğdır Province in Türkiye. For this purpose, 60 brain tissue samples of sheep were collected within the lambing seasons of 2023 in Iğdır, Türkiye. The DNA extraction was performed using the PureLink™ Genomic DNA Mini Kit from brain samples. The PCR was performed with the appropriate primers from the obtained DNA samples. T. gondii was found in the brain (16.6%) samples of aborted sheep fetuses. According to the present study, T. gondii infection can be one of the causes of fetus abortion of sheep in Iğdır province, Türkiye. This result emphasizes the need for vigilance and preventive measures in managing this potential public and animal health concerns

Molecular Survey of the Toxoplasma gondii and Neospora caninum in brain tissue of aborted fetuses of Morkaraman sheep in Muş, Türkiye

Toxoplasma gondii and Neospora caninum are obligate intracellular protozoan parasites that can affect different warm-blooded species worldwide. In this study, it was aimed to detect T. gondii and N. caninum using PCR method in brain tissues of aborted sheep fetuses. Brain specimens were collected from 50 Morkaraman sheep fetuses that had undergone abortion at various stages of pregnancy, within the lambing seasons of 2023 in Muş. Approximately 1 cm³ of brain tissue from the right cerebral hemisphere was excised and subsequently frozen at -20°C for DNA extraction. DNA extraction and PCR amplification were then performed. As a result of this study, 11 (22%) of 50 brain tissues were positive. All brain samples examined in this study were negative for Neospora caninum. Based on the results of this study, it is possible to say that T. gondii is an important abortion agent in sheep in this region. Although N. caninum was not detected in this study, larger scale studies are recommended. Moreover, this study provides important information to breeders and veterinarians in the field in the evaluation and management of abortion

The efficacy of chloroquine treatment against naturally occuring Giardia duodenalis infection in lambs

Objective. The aim of the present study was to determine the efficacy of Chloroquine (Cq), an antimalarial medicine, administered at a dose of 2.5 mg/kg, orally, during 5 consecutive days, in Sakiz and Merino lambs naturally infected with Giardia duodenalis. Materials and methods. To this extent weaned 10 weeks of aged lambs were enrolled and randomly assigned into two groups based on treatment (group C, n=18 lambs treated with Cq) and placebo (group P, n=8 untreated control animals). Diagnosis was based on detection of trophozoit and/or cysts on fecal flotation. Cyst count per gram feces (days 0, 3, 7 and 10) was analyzed among groups. Results. During the trial, regarding the efficacy of Cq on days 3., 7., and 10. There was significant (p<0.01) reduction in cyst excretion; whereas evaluation of mean geometric cyst excretion revealed 100% reduction. Conclusions. There was a very high (100%) reduction in cyst excretion in the Cq treatment group compared to the positive untreated control group on days 3, 7 and 10, resulting in a significant (p<0.001) reduction, making Cq, safety, and thus highly effective treatment option of lambs with naturally occuring giardiasis. Objetivo. El objetivo del presente estudio fue determinar la eficacia de la cloroquina, un medicamento contra la malaria, administrado a una dosis de 2.5 mg/kg, por vía oral, durante 5 días consecutivos, en Corderos Sakiz y Merino infectados naturalmente con Giardia duodenalis. Materiales y metodos. En esta medida destetados 10 semanas de edad fueron incluidos y asignados al azar en dos grupos sobre la base del tratamiento (grupo C, n = 18 corderos tratados con cloroquina) y placebo (grupo P, n = 8 animales de control no tratados). El diagnóstico se basa en la detección de trophozoit y/o quistes en la flotación fecal. Quiste recuento por gramo de heces (día 0, 3, 7 y 10) se analizó entre los grupos. Resultados. Respecto a la eficacia de la cloroquina en los días 3, 7 y 10 existió una reducción significativa (p<0.01) en la excreción de quistes; mientras que la evaluación de la media de la excreción Rev.MVZ Córdoba 21(2):5328-5335, 2016. ISSN: 0122-0268 ORIGINAL 5329 de quistes reveló una reducción del 100%. Conclusiones. Durante el estudio hubo una reducción del 100% en la excreción de quistes en el grupo de tratamiento con cloroquina en comparación con el grupo control no tratado positivo en los días 3, 7 y 10, lo que resulta en una disminución significativa (p<0.001) por lo que la cloroquina razonablemente es una opción de tratamiento por el costo, seguridad, y por lo tanto muy eficaz de corderos infectados naturalmente con giardiasis

Genetic characterization of Varroa destructor (Family: Varroidae) prevalent in honeybees (Apis mellifera) in the province of Aydin in Turkey

The aim of the present study was to identify the haplotypes of the Varroa destructor mite which infects honeybees in the province of Aydın in Turkey, using two different modified techniques for the mitochondrial Cox1 gene of the mite. In order to confirm the haplotype, two primers differing in their sequence i.e. (ADA 01) as forward primer 5′-TACAAAGAGGGAAGAAGCAGCC-3′ and reverse primer 5′- GCCCCTATTCTTAATACATAGTGAAAATG-3′ and (ADA 02) with COXF primer [5′GG(A/G)GG(A/T)GA(C/T)CC(A/T)ATT(C/T)T(A/T)TATCAAC3′] and COXRa primer [5′GG(A/T)GACCTGT(A/TA(A/T)AATAGCAAATAC3′], were selected. Amplified DNA 376 bp in size was acquired using (ADA 01) forward primer 5′- TACAAAGAGGGAAGAAGCAGCC-3′ and reverse primer 5′- GCCCCTATTCTTAATACATAGTGAAAATG-3′. SacI restriction enzyme was applied to the amplified products; however, this restriction enzyme did not cut the DNA. Amplified DNA, 570 bp in size was obtained using (ADA 02) COXF primer [5’GG(A/G)GG(A/T)GA(C/T)CC(A/T)ATT(C/T)T(A/T)TATCAAC3’] COXRa and [5’GG(A/T)GACCTGT(A/TA(A/T)AATAGCAAATAC3’]. XhoI and SacI restriction enzymes were used for the amplified products. Although, the SacI restriction enzyme did not cut the DNA, the XhoI restriction enzyme cut the amplified DNA into two fragments (bands), with the sizes of 270 and 300 bp two bands 270 and 300 bp. While comparing the results, these bands were found specific for Korean haplotype of V. destructor. In conclusion, all of the 200 samples of V. destructor examined in this study were identified to be the Korean haplotype