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Analysis of cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) gene related to neonatal isoerythrolysis in stray cats of Izmir, Turkey [Türkiye, İzmir sokak kedilerinde neonatal izoeritrolizisle ilişkili sitidin monofosfat-N-asetilnöraminik asit hidroksilaz (CMAH) geninin analizi]
(Veteriner Fakultesi Dergisi, 2016) Can H.; Atalay Şahar E.; Döşkaya M.; Özdemir H.G.; Caner A.; Değirmenci Döşkaya A.; Gürüz Y.; Ün C.
Neonatal isoerythrolysis is a life threatening disease in new born cats. It occurs when type A or type AB kittens are born from a type B queen (female cat). A homozygous 18 bp insertion located in cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) gene has been detected in type B cats, causing production of inactive CMAH enzyme. Currently, molecular methods are being used to determine type B blood in female cats, which can help prevent neonatal isoerythrolysis in kittens. These molecular assays target the presence of 18 bp insertion in CMAH gene. In this study, we aimed to analyze the potential of neonatal isoerythrolysis among stray cats of İzmir, Turkey using PCR detecting the 18 bp insertion in CMAH gene. During the study, we analyzed 793 cats’ blood sample for the presence of 18 bp insertion in CMAH gene. Three cats known to have blood types A, B, and AB were used as control in PCR. According to the PCR results, blood type A control cat displayed a 175 bp product indicating a homozygous type A cat while blood type control B cat showed a 193 bp product in CMAH gene (with 18 bp insertion) indicating a homozygous type B cat. Interestingly, blood type AB control cat showed a heterozygous pattern for CMAH gene, in which three different bands (175 bp like that of type A, 193 bp product for type B, and the third unique band with approximately 240 bp size) were detected. Among 793 stray cats of İzmir, 791 were homozygous for CMAH gene with 175 bp band size (99.7%). The remaining two stray cats showed heterozygous band pattern like blood type AB cat (0.12%). Overall, 175 bp band displaying type A cats are prevalent contrary to the two cats that have type AB pattern and non-existence of homozygous type B cats. These results show that the potential of neonatal isoerythrolysis in stray cats of İzmir is minimal. Future studies are required to scrutinize the reason(s) for non-existence of type B cats in İzmir and presence of unique band in blood type AB. © 2016, Veteriner Fakultesi Dergisi. All rights reserved.
Arugula and pre-converted arugula extract as natural Nitrate/Nitrite sources for heat-treated sausages
(IOP Publishing Ltd, 2021) Can H.; Sari B.; Kavuşan H.S.; Serdaro?lu M.
The aim of this study was to investigate using arugula or pre-converted extracts as nitrite alternatives in heat-treated fermented sausages. Sausages with nitrite, 150 mg/kg of NaNO2, 1.2% arugula extract, and 1.5% pre-converted arugula extract were formulated. Natural nitrate sources added resulted in significantly lower oxidation content compared to negative control groups at initial storage. The addition of natural nitrate sources influenced colour, pigments, and conversion rate of sausages. Pre-converted arugula extract showed little effect on the residual nitrite content. The result of colour, oxidation, and nitrite analysis suggest pre-converted arugula is a potential nitrite replacer, but arugula as a nitrate source is limited to provide the functions of nitrite. © Published under licence by IOP Publishing Ltd.
Characteristics of various cell types in primary insect cell culture from larval ovaries of silkworm (Bombyx mori L.)
(Pakistan Agricultural Scientists Forum, 2019) Soya S.; Can H.
As insect cell lines can be used in baculovirus expression systems for producing recombinant proteins, there is a need to establish new cell lines. Insect cell lines are thought to be useful in production of humanized recombinant proteins. Hence, we aimed to establish a culture from larval ovaries of Bombyx mori to investigate the cell characteristics and growth properties. Two sets of primary cell cultures were established from ovaries of fifth instar larvae. Characterization of cell line was carried out by comparing the PCR products of cell line and silkworms from COI (cytochrome c oxidase subunit I) specific primers. In the first set of cultures, the culture population was found to be heterogeneous and cell sizes were variable. Fibroblastic, epithelial-like, spindle-like, and round cells were observed after 7 th day. Interestingly, contractions were observed in muscle-like cells at the end of first month, and stayed stable until the fourth month. At the beginning of the fourth month, because cell death increased significantly, cell concentrations decreased. In the second set of cultures, it was determined that doubling time was about 72 h with high viability and confluency of cells were higher in early passages. PCR which was conducted for characterization of cells resulted in a 710 bp product in each cell line and silkworms. In conclusion, growth rate detected in this study was observed to be compatible with literature. Also, it was thought that if primary culture is supported with growth stimulators, growth rate can be more accelerated and thi s primary culture can be used more efficiently in recombinant protein production studies. © 2019, Pakistan Agricultural Scientists Forum. All rights reserved.
Collaborative effect of fat reduction and ?-tocopherol incorporation on oxidative stability in beef sausages
(Institute of Physics Publishing, 2019) Nacak B.; Kavusan H.S.; Sari B.; Can H.; Serdaroglu M.
This study focuses on the changes in oxidative stability and sensory properties of reduced fat and/or ?-tocopherol incorporated sausages during storage at 4°C for 3 months. In order to examine these changes, sausages were formulated with 20% fat and 20% fat+200 ppm ?-tocopherol, coded as C20A0 and C20A1, respectively. Sausages formulated with 10% fat (C10A0) and 10% fat+200 ppm ?-tocopherol (C10A0) were low fat sausages. Reduction of fat by 50% or adding ?-tocopherol initially increased the peroxide values of sausages, but at the end of storage, conversely, reduction of fat and ?-tocopherol addition retarded lipid peroxidation as well as malonaldehyde generation (p<0.05). The highest thiobarbituric acid reactive substance values were recorded for C20A0 sausages initially and at the end of the storage (p<0.05). Even though thiobarbituric acid reactive substance values of sausages with 20% fat were higher, the initial rancid taste of all sausages were similar at month 0, but differences in the rancid taste of sausages began to be revealed with increasing storage time (p<0.05). No significant differences were found in the general acceptability of all the sausages during the 3-month storage period (p>0.05). © Published under licence by IOP Publishing Ltd.
Comparison of the effects of Artemisia vulgaris and Artemisia absinthium growing in western Anatolia against trichinellosis (Trichinella spiralis) in rats
(2008) Caner A.; Döşkaya M.; Degirmenci A.; Can H.; Baykan S.; Üner A.; Başdemir G.; Zeybek U.; Gürüz Y.
Trichinellosis often causing diarrhea and more rarely fever, periorbital edema and myositis in human, is commonly treated with benzimidazole derivatives. The Artemisia genus has been found to be effective against a variety of parasites. In the present study, the efficacy against trichinellosis (Trichinella spiralis) of Artemisia vulgaris and Artemisia absinthium was examined for the first time in rats. The results of trichinoscopy and artificial digestion, during the enteral (adult) phase of the illness show that 300 mg/kg doses of methanol extracts of the aerial parts of A. vulgaris and A. absinthium reduced the larval rate by 75.6% and 63.5% in tongue, 53.4% and 37.7% in diaphragm, 67.8% and 46.2% in quadriceps, and 66.7% and 60.5% in biceps-triceps muscles of rats, respectively. Furthermore, during the parenteral (encapsulated larvae) phase, 600 mg/kg doses of A. vulgaris and A. absinthium extracts decreased the larval rate by 66.4% and 59.9% in tongue, 57.4% and 50.0% in diaphragm, 47.6% and 43.7% in quadriceps, 60.2% and 46.4% in biceps-triceps muscles of rats, respectively. Analysis of antibody also showed that A. vulgaris significantly reduced the antibody response (P < 0.05) during the enteral and parenteral phases. Thus, the results of the present study revealed that A. vulgaris could be an alternative drug against trichinellosis. © 2008 Elsevier Inc. All rights reserved.
[Cryopreservation of Toxoplasma gondii tachyzoites and tissue cysts]. [Toxoplasma gondii Takizoit ve Doku Kistlerinin Kriyoprezervasyonu.]
(2013) Döşkaya M.; Caner A.; Can H.; Iz S.G.; Degirmenci A.; Gürüz A.Y.
Toxoplasma gondii tachyzoites and tissue cysts are largely used for developing diagnostic assays, vaccines and in drug research as well as biochemical and molecular structure studies. Continuous passaging of tachyzoites or tissue cysts in animal models encounter ethical and economical problems and it is a time consuming procedure. Cryopreservation of tachyzoites and tissue cysts and revitalization of cryopreserved samples whenever needed, can decrease the economical loss, ethical problems and labour. In the present article, production of tachyzoites and tissue cysts in mice, preparation of samples for cryopreservation, cryopreservation of tachyzoites and tissue cysts, defrosting of cryopreserved samples and reinoculation to mice have been described in detail.
[Demonstration of Cryptosporidium parvum in immune suppressed rats using nested PCR] [İmmun Sistemi Baskılanmış Sıçanlarda Cryptosporidium parvum'un Nested PZR Yöntemi ile Gösterilmesi]
(2013) Can H.; Caner A.; Döşkaya M.; Değirmenci A.; Karaçali S.; Gürüz Y.; Uner A.
[No abstract available]
Detection of echinococcus granulosus and echinococcus multilocularis in cyst samples using a novel single tube multiplex real-time polymerase chain reaction [Kist Örneklerinde Yeni Bir Tek Tüp Multipleks Gerçek Zamanli Polimeraz Zincir Reaksiyonu He Echinococcus granulosus ve Echinococcus multilocularis' in Saptanmasi]
(Ankara Microbiology Society, 2016) Can H.; Inceboz T.; Caner A.; Atalay Şahar E.; Karakavuk M.; Döşkaya M.; Çelebi F.; Degirmenci Döşkaya A.; Gülçeiz S.; Gürüz Y.; Korkmaz M.
Cystic echinococcosis (CE) and alveolar echinococcosis (AE) caused by Echinococcus granulosus and Echinococcus multilocularis, respectively, are important helminthic diseases worldwide as well as in our country. Epidemiological studies conducted in Turkey showed that the prevalence of CE is 291-585/100.000. It has also been showed that the seroprevalence of AE is 3.5%. For the diagnosis of CE and AE, radiological (ultrasonography, computed tomography, magnetic resonance) and serological methods, in addition to clinical findings, are being used. The definitive diagnosis relies on pathological examination When the hydatid cysts are sterile or does not contain protoscolex, problems may occur during pathological discrimination of E.granulosus and E.multilocularis species. In this study, we aimed to develop a novel multiplex real-time polymerase chain reaction (M-RT-PCR) targeting mitochondrial 12S rRNA gene of E.granulosus and E.multilocularis using Echi S (5'-TTTATGAATATTCTGACCCTGACAT-3') and Echi A (5'-GGTCTTAACTCMCTCATGCAG-3') primers and three different probes; Anchor Ech (5'-GTTTGCCACCTCGATGTTGACTTAG-fluoroscein-3'), Granulosus (5'-LC640-CTMGGTTTTGGTGTAGTMTTGATATTTT-phosphate-3') and Multilocularis (5'-LC705-CTGTGATCTTGGTGTAGTAGTTGAGATT-phosphate-3') that will enable the diagnosis of CE and AE in same assay. During M-RTR-PCR, plasmids containing E.granulosus (GenBank: AF297617.1) and E.multilocularis (GenBank: NC-000928.2) mitochondrial 12S rRNA regions were used as positive controls. Cysts samples of patients which were pathologically confirmed to be CE (n: 10) and AE (n: 15) and healthy human DNA samples (n: 25) as negative control as well as DNA samples of 12 different parasites (Taenia saginata, Hymenolepis nana, Trichuris trichiura, Fasciola hepatica, Enterobius vermicularis, Toxoplasma gondii, Pneumocystis jirovecii, Trichomonas vaginalis, Cryptosporidium hominis, Strongyloides stercoralis, Plasmodium falciparum, Plasmodium vivax) were used to develop M-RT-PCR. E.granulosus and E.multilocularis control plasmids were constructed to detect analytic sensitivity of the test using TOPO cloning. Positive control plasmids were diluted to determine analytical sensitivity and specificity by distilled water at 106-105-104-103-102-101-1 plasmid copy of dilution in each reaction. According to the results, analytical sensitivity of the assay for E.granulosus and E.multilocularis was 1 copy plasmid/µl reaction. The non-existence of cross reactivity with 12 different parasites' DNA samples showed the analytical specificity of the assay. Displaying Echinococcus DNA in cyst samples among 25 patients and species discrimination as well as non-existence of cross reactivity with human DNA samples showed that the clinical sensitivity and specificity of the assay were 100%. As a result, the M-RT-PCR developed in the present study provided a sensitive, specific, rapid, and reliable method in the diagnosis of echinococcosis and the discrimination of E.granulosus and E.multilocularis from cyst samples.
Detection of Pneumocystis in the nasal swabs of immune-suppressed rats by use of PCR and microscopy.
(2013) Can H.; Caner A.; Döşkaya M.; Degirmenci A.; Karaçali S.; Polat C.; Gürüz Y.; Uner A.
Detection of Pneumocystis jiroveci colonization in lungs or oral samples due to high sensitivity of PCR methods results in undue treatment of patients without any symptoms of Pneumocystis pneumonia. The aim of the present study is to demonstrate Pneumocystis carinii in rats, immune suppressed by oral and subcutaneous administration of dexamethasone. Blood, oral, nasal and eye swabs were collected prior to immune suppression and 2, 6, 12 weeks after administration of dexamethasone. Also, samples were collected from lung, heart, liver, kidney, diaphragm, brain, spleen, tongue, muscle, eye, intestine, and feces. Cysts and trophozoites were investigated in stained slides and MSG gene was detected by PCR. The results showed that weight loss is significantly higher in rats administered oral dexamethasone (P<0.05). Microscopy was positive only in lungs of rats orally administered dexamethasone. PCR was positive in lungs and oral swabs of rats prior to the administration of dexamethasone. After the administration of dexamethasone, the MSG gene was detected in oral swabs, lungs, spleen, kidney and (for the first time) in nasal swabs. PCR was positive in nasal swabs during the second and sixth weeks of oral and subcutaneous administration of dexamethasone, respectively. Presence of P. jiroveci in nasopharyngeal aspirate, oropharyngeal wash, oral swab, induced sputum or BAL, and absence in nasal swab in a patient without symptoms of PCP may support clinician's decision regarding colonization. Overall, detection of P. carinii in nasal swabs of rats by PCR demonstrated that nasal sampling can be used for the diagnosis of Pneumocystis pneumonia.
Detection of Toxoplasma gondii in a Eurasian Badger (Meles meles) Living in Wildlife Areas of Izmir, Turkey
(NLM (Medline), 2018) Karakavuk M.; Aldemir D.; Atalay Şahar E.; Can H.; Özdemir H.G.; Değirmenci Döşkaya A.; Gürüz A.Y.; Döşkaya M.
Toxoplasma gondii is an obligatory intracellular protozoon parasite that causes toxoplasmosis in humans and all warm-blooded animals. In this study, we aimed to investigate the presence of T. gondii DNA in a Eurasian badger (Meles meles) that was found dead in the wildlife area of Izmir. According to the results of real time polymerase chain reaction, T. gondii REP gene was found to be positive in the Eurasian badger brain homogenate. In conclusion, Eurasian badger, a known carnivore, can be a potential source of toxoplasmosis in the natural settings of İzmir, Turkey.
Diagnosis and Species Discrimination of Positive Malaria Samples by Real-Time Polymerase Chain Reaction
(2016) Atalay Şahar E.; Karakavuk M.; Can H.; Nergiz Ş.; Gül K.; Değirmenci Döşkaya A.; Gürüz Y.; Döşkaya M.
OBJECTIVE: Malaria is an important tropical disease that is detected in 198 million people and causes 367-755 thousand deaths annually. Recently, the real-time Polymerase Chain Reaction (PCR) technique has enabled quick determination of Plasmodium spp. and species identification in the same assay with a low contamination risk. In the present study, we aimed to use real-time PCR targeting the 18S rRNA gene to diagnose Plasmodium spp. and perform species identification.METHODS: DNA samples of 15 patients with malaria (14 caused by P. vivax, 1 caused by P. falciparum) confirmed by microscopy as well as positive control plasmids were used. As the negative control, DNA samples of 15 individuals without malaria were used.RESULTS: According to the results of real-time PCR, samples of 15 patients with malaria were found to be positive for Plasmodium spp. Melting curve analysis showed that 14 of them were P. vivax and the remaining was P. falciparum. In addition, mixed infection with P. falciparum and P. vivax was successfully detected by real-time PCR when DNA of P. falciparum- and P. vivax-positive samples was experimentally mixed.CONCLUSION: The present study showed that real-time PCR can be useful in the diagnosis and species identification of Plasmodium spp. as well as the detection of mixed infections in addition to microscopy in Turkey.
Diagnostic value of a Rec-ELISA Using toxoplasma gondii recombinant SporoSAG, BAG1, and GRA1 proteins in murine models infected orally with tissue cysts and oocysts
(Public Library of Science, 2014) Döşkaya M.; Caner A.; Can H.; Iz S.G.; Gedik Y.; Döşkaya A.D.; Kalantari-Dehaghi M.; Gürüz Y.
Toxoplasma gondii causes congenital toxoplasmosis in newborns resulting with fetal anomalies. Determining the initiation time of infection is very important for pregnant women and current serological assays have drawbacks in distinguishing the recently acute toxoplasmosis. Diagnosis of recently acute infection may be improved by using stage specific antigens in serological assays. In the present study, the diagnostic value of sporozoite specific SporoSAG, bradyzoite specific BAG1 proteins and GRA1 protein expressed by all forms of the parasite have been evaluated ELISA using sera systematically collected from mice administered orally with tissue cyst and oocysts. The anti-SporoSAG IgM antibodies in sera obtained from mice infected with oocysts peaked significantly at days 1, 10, and 15 (P<0.01). The anti-BAG1 IgM antibodies in sera obtained from mice infected with tissue cysts peaked significantly at days 15, 40, and 120 (P<0.05). The anti-GRA1 IgM antibodies in sera obtained from mice infected with oocysts peaked significantly at days 2, 10, and 40 (P<0.01). The anti- GRA1 IgM antibodies in sera obtained from mice infected with tissue cysts peaked significantly only at day 40 (P<0.05). The anti-SporoSAG, anti-BAG1, and anti-GRA1 IgG titers of mice showed significant increases at day 40 (P<0.05) and decrement started for only anti-GRA1 IgG at day 120. The presence of anti-SporoSAG IgM and IgG antibodies can be interpreted as recently acute infection between days 10-40 because IgM decreases at day 40. Similarly, presence of anti-BAG1 IgM and absence of IgG can be evaluated as a recently acute infection that occurred 40 days before because IgG peaks at day 40. A peak in anti-GRA1 antibody level at first testing and reduction in consecutive sample can be considered as an infection approximately around day 40 or prior. Overall, recombinant SporoSAG, BAG1 and GRA1 proteins can be accepted as valuable diagnostic markers of recently acute toxoplasmosis. Copyright: © 2014 Döşkaya et al.
Elimination of phosphate in restructured turkey steaks by the addition of eggshell calcium powder and low methoxyl pectin
(IOP Publishing Ltd, 2021) Kavuşan H.S.; Yüncü O.; Can H.; Serdaro?lu M.
This study was carried out to assess the effects of eggshell calcium powder (ESCP) and/or low methoxyl pectin (LMP) as phosphate replacers on the quality parameters of restructured turkey steaks. ESCP, 0.25% or 0.50%, was added to formulation alone or in combination with 0.25% LMP in powder and gel forms. The pH increased with the addition of ESCP. Soluble protein content, water holding capacity, and cooking yield were higher in steaks formulated with ESCP+LMP gel compared to control steaks containing phosphate. Hardness of steaks was decreased by the addition of ESCP and pectin. Pectin in powder form negatively affected the preference of panelists. Oxidation in phosphate-free steaks was more pronounced than in other treatments. The results showed that the binding properties of phosphate could be achieved by using ESCP or ESCP+LMP gel. © Published under licence by IOP Publishing Ltd.
Genetic characterization of Toxoplasma gondii isolates and toxoplasmosis seroprevalence in stray cats of Izmir, Turkey
(2014) Can H.; Döşkaya M.; Ajzenberg D.; Özdemir H.G.; Caner A.; Iz S.G.; Döşkaya A.D.; Atalay E.; Çetinkaya Ç.; Ürgen S.; Karaçali S.; Ün C.; Dardé M.-L.; Gürüz Y.
Currently, some Toxoplasma gondii genotypes are being associated with serious clinical presentations. A recent report showing the Africa 1 genotype in two local congenital toxoplasmosis cases acquired in Turkey formed the basis of this study because atypical Africa 1 genotype is most frequently detected in animals and patients from sub-Saharan Africa. Since stray cats are considered as the linkage between wild life and urban life in T. gondii transmission, the present study aimed to isolate and characterize T. gondii strains circulating in stray cats of İzmir (Western Turkey). A secondary objective was to determine toxoplasmosis seroprevalence in this cat population. Tissues obtained from 100 deceased stray cats were bioassayed and isolated strains were genotyped using 15 microsatellite markers. In addition, toxoplasmosis seroprevalence was analyzed in 1121 cat sera collected from several large veterinary clinics in İzmir. Among the 22 isolates, 19 were Type II (86.3%), two were Type III (9%) and one was Africa 1 genotype (4.5%). The overall seropositivity rates in cats were 42-48% and 33.4-34.4% according to IFA and ELISA, respectively. Seroprevalence in deceased cats was significantly higher than in healthy cats (P = 0.0033). Finding both the major clonal Type II lineage together with the Type III lineage also found in Middle East, and an atypical genotype, Africa 1 appears consistent with the specific geographic location of Turkey between three continents and raises the possibility of transportation of these strains between continents through trade routes or long distance migratory birds. In addition, the first large study of toxoplasma seroprevalence in a stray cat population was also reported. The relatively high seropositivity rates and the variety of T. gondii genotypes confirm the local stray cat population as a risk factor for human toxoplasmosis in İzmir. © 2014 Can et al.
Immunogenic multistage recombinant protein vaccine confers partial protection against experimental toxoplasmosis mimicking natural infection in murine model
(Elsevier B.V., 2016) Gedik Y.; Gülçe Iz S.; Can H.; De?irmenci Döşkaya A.; Ismet Delilo?lu Gürhan S.; Gürüz Y.; Döşkaya M.
Toxoplasma gondii is a protozoan parasite that can infect warm-blooded animals including humans. Vaccination studies mostly use tachyzoite specific proteins however in natural route of infection, toxoplasmosis initiates with tissue cysts (bradyzoites) or oocysts (sporozoites) and thereafter stage conversion takes place where the tachyzoites take action and cause acute infection continues with tachyzoites. Despite this knowledge, challenging models used in the vaccination studies prefer administration of tissue cyst forming strains intraperitoneally or subcutaneously instead of oral administration which is the natural route of infection. In the present study, a multivalent adjuvanted recombinant protein vaccine that contains bradyzoite specific BAG1 and tachyzoite specific GRA1 protein and controls were administered to female Swiss Webster outbred mice. Humoral and cellular immune responses were analyzed by Rec-ELISA, Western blot, and flow cytometry. Mice were infected orally with T. gondii PRU strain tissue cysts using feeding needle to mimic the natural route of infection. 40 days after challenging microscopy and Real Time PCR were performed to determine the protection level. Analysis of sera obtained from vaccinated mice showed strong anti-BAG1 and anti-GRA1 IgG responses. The IgG2a response was significantly higher (P < 0.0001) and the ratio of CD8 + T lymphocytes secreting IFN-? almost doubled compared to PBS control which are indicative of protection against toxoplasmosis. The amount of tissue cysts in vaccinated group was reduced 10.5% compared to control group. To generate a protective vaccine against toxoplasmosis, multistage vaccines and usage of challenging models mimicking natural route of infection are critical cornerstones. In this study, we generated a BAG1 and GRA1 multistage vaccine that induced strong immune response in which the protection was not at anticipated level. In addition, the murine model was orally challenged with tissue cysts to mimic natural route of infection. © 2015 Published by Elsevier Ltd.
The importance of the contribution of rapid test, serological and molecular methods in the diagnosis of two imported malaria cases with atypical microscopy [Mikroskopide Atipik Gorunumlu Dis Kaynakli Iki Sitma Olgusunda Hizli Test, Serolojik ve Molekuler Yontemlerin Taniya Katkisinin Onemi]
(Ankara Microbiology Society, 2017) Zorbozan O.; Pullukcu H.; Sahar E.A.; Karakavuk M.; Can H.; Tunali V.; Doskaya M.; Turgay N.; Toz S.; Ozbilgin A.
Malaria is a widespread and life-threatening disease in tropical and subtropical regions. In patients with typical clinical symptoms, malaria is considered as a preliminary diagnosis if there is a travel history to malaria-endemic areas. The basis of the laboratory diagnosis of malaria is the microscopic examination of Giemsa stained smears. On the other hand, the diagnosis and differentiation of Plasmodium species with microscopic examination may have some difficulties. In the first case, adifferent appearance from the classical Plasmodium vivax erythrocytic forms in infected erythrocytes were detected in 1% of all erythrocytes in thin smear blood preparations of a 26-year-old male with complaints of fever and chills and a story of travel to Nigeria. It was observed that parasitic nuclei were not prominent, and were located in the cytoplasm irregularly as chromatin or dye particles, nucleus fragments similar to Schiiffner's granules in the form of scattered and granular spots were present in some erythrocytes, the cytoplasm of some Plasmodium erythrocytic forms were irregular and nuclei were not seen. There were no Schiiffner's granules in any of the infected erythrocytes. PMvax was detected by the rapid diagnostic test (OptiMAL, DiaMed GmbH, Switzerland), which searches for the antigens of Plasmodium species, in the peripheral blood sample of the patients. The P.vlvax 18S rRNA gene was also detected by the multiplex real-time polymerase chain reaction. Antibodies against Plasmodium species were searched by using the Pan Malaria Antibody CELISA (CeLLabs Pty Ltd, Brookvale, Australia) kit in the patient's serum sample and the optical density (OD) value of the patient sample was measured five times the OD value of the positive control. In the second case, adifferent appearance from the classical P.faldparum erythrocytic forms in infected erythrocytes were detected in 12% of all erythrocytes in thin smear blood preparations of a 31-year-old male who has been suffering from persistent fever, severe headache, pain in the eyes and was known to be working in Nigeria. It was observed that some Plasmodium trophozoites have 1 /3 of the size of erythrocytes such as P.vivax and have non-granular cytoplasm, some erythrocytic forms were round and the nucleus and cytoplasm were hardly distinguished, some of them were seen as crescent and close to the nucleus of the cytoplasm and some erythrocytic forms had characteristically a single nucleus and a scattered cytoplasm, similar to mature trophozoites of P.vivax. Although the Plasmodium young trophozoites were similar to Rvtvax in means of magnitude, the forms in which the nude adhered to the erythrocyte wall were common. There were no Rfalciparum gametocyte forms. Rfalciparum like young trophozoite was observedonly in one of the four smears. P.falciparum was detected by the commercial rapid diagnostic test and Rfalciparum 18S rRNA gene was also detected by the multiplex real-time polymerase chain reaction. Antibody formation against Plasmodium species was not detected in the ELISA test. In these case reports, the importance of the support of rapid diagnostic tests, serological and molecular methods to microscopic diagnosis and species determination of two imported malaria cases were demonstrated.
In vitro anti-leishmanial activity of Sarcopoterium spinosum against Leishmania tropica [Sarcopoterium spinosum’un Leishmania tropica’ya karşı in vitro anti-leishmanial etkisi]
(Chartered Inst. of Building Services Engineers, 2018) Can H.; Kayalar H.; Bozkurt B.; Can Ş.; Döşkaya M.; Töz S.
Complex clinical symptoms such as ulcerative skin lesions, destructive mucosal inflammation, and disseminated visceral infection can reveal in leishmaniasis. The conventional drugs are toxic and expensive. In addition, patients receive a long treatment with these drugs which have adverse effects and unfortunately there are some limitations during the treatment. The aim of this study is to investigate the in vitro anti-leishmanial activities of four different extracts of Sarcopoterium spinosum against Leishmania tropica. Initially, different concentrations of ethanol, methanol, n-hexane, and water extracts of S. spinosum were incubated with L. tropica promastigotes. After 72 hours of incubation, the growth of L. tropica promastigotes was significantly inhibited and the percentage of inhibition ranged between 42.8 and 100 %. Among these extracts, the most efficient growth inhibition (100 %) was obtained with methanol extract (at a dose of 50 µg/ml). In conclusion, S. spinosum may be a potential source for the development of novel therapeutic agents to treat L. tropica infection. © 2018, Chartered Inst. of Building Services Engineers. All rights reserved.
Investigation of Freshness and Oxidative Changes of Fried Sardine Marinades with Barberry or Acorn Extracts
(Josip Juraj Strossmayer University of Osijek Faculty of Food Technology, 2024) Kavuşan H.S.; Yüncü-Boyaci Ö.; Can H.; Düz İ.G.; Karataş S.; Meltem S.
This study was carried out to investigate the effects of using barberry or acorn extracts in fried sardine marinade formulations. In line with this purpose, sardines were eviscerated, washed, and filleted. Fillets were marinated with a solution containing 4% salt, 1% vinegar, and 0.2% potassium sorbate. After that, four marinated batches were made, namely C (no antioxidant added), B (200 ppm BHT added), BE (barberry extract added-200 ppm gallic acid equivalent (GAE)), and AE (acorn extract added-200 ppm gallic acid equivalent). Marination uptake, chemical composition, pH, colour, oxidation (peroxide value and lipid oxidation), trimethylamine (TMA-N), and sensory analyses were performed at +4 ? for 28 days. Acorn extract exhibited higher total phenolic content and pH value than barberry extract. Both barberry and acorn extract contain various bioactive components, especially phenolic acids, flavonoids, and alkaloids. The incorporation of antioxidants did not alter the chemical composition, initial pH and marinade uptake. Barberry extract was responsible for the lowest oxidation on the 14th day of storage. TBARS and TMA-N values of all extract added treatments were within the acceptable limits. Only flavour and general acceptability scores of AE were lower. All treatments were preferred at the same level except for AE treatment. © 2024 by the author(s).
Investigation of the role of stray cats for transmission of toxoplasmosis to humans and animals living in Izmir, Turkey
(Journal of Infection in Developing Countries, 2021) Karakavuk M.; Can H.; Selim N.; Yesilsiraz B.; Atli E.; Sahar E.A.; Yalçin M.
Introduction: Toxoplasma gondii is a protozoan parasite that has a widespread distribution among mammalians and birds. One of the reasons for the high prevalence may be due to ingesting oocyst disseminated by stray cats' feces. In Turkey, most of the citizens are closely associated with stray cats and they love to pet and feed them on the streets. In this study, we aimed to determine the prevalence of T. gondii DNA in feces of stray cats living in Izmir, Turkey in order to identify the transmission potential to humans and other animals. Methodology: Feces and blood samples of 465 stray cats were investigated for the presence of T. gondii oocysts by microscopy and for the presence of T. gondii DNA by two real time PCR methods. Furthermore, serum samples were analyzed for anti-T. gondii IgG antibodies using an ELISA. Results: Oocysts were detected in 0.43% of the stray cats by microscopy. T. gondii DNA was detected in 14.37% of the stray cats' feces samples. The seroprevalence rate was 37.84%. In the feces and/or blood PCR positive group, 35.89% of them were seropositive. Among the 176 seropositive cats, T. gondii DNA was detected in feces of 27 cats (15.34%). Conclusions: This study first time showed the inter relation of T. gondii DNA in feces and blood samples and seropositivity. In sum, over 14% of the stray cats living outdoor may have an important role in transmission of toxoplasmosis to humans in Izmir as well as to other animals. © 2021 Journal of Infection in Developing Countries. All rights reserved.
Molecular characterization of cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) gene and frequency of blood types in stray cats of İzmir, Turkey
(BioMed Central Ltd, 2021) Can H.; Erkunt Alak S.; Köseoğlu A.E.; Şahar U.; Bostanbaş B.; Baydarlı S.; Ün C.
Background: Cytidine monophospho-n-acetylneuraminic acid hydroxylase (CMAH) gene associated with blood groups in cats encodes CMAH enzyme that converts Neu5Ac to Neu5Gc. Although variations in CMAH gene of pedigree cats have been revealed, the presence/lack of them in non-pedigree stray cats is unknown. Therefore, the present study aimed to investigate the variations in CMAH gene and the quantity of Neu5Ac and Neu5Gc on erythrocytes of non-pedigree stray cats (n:12) living in İzmir, Turkey. Also, the frequency of blood types was determined in 76 stray cats including 12 cats that were used for CMAH and Neu5A/Neu5Gc analysis. Results: In total, 14 SNPs were detected in 5’UTR as well as in exon 2, 4, 9, 10, 11 and 12 of CMAH gene. Among these SNPs, -495 C > T in 5’UTR was detected for the first time as heterozygous in type A and AB cats, and homozygous and heterozygous in type B cats. The remaining 13 that have been detected in previous studies were also found as homozygous or heterozygous. Both Neu5Gc and Neu5Ac were detected in type A and AB cats. In type B cats, only Neu5Ac was detected. Among two type AB cats, the level of Neu5Ac was found higher in cat carrying heterozygous form (T/C) of 1392T > C. The prevalence of type B cats (67.1 %) was higher than others. Conclusions: The presence of a new SNP as well as previous SNPs indicates that more variations can be found in stray cats with a more comprehensive study in the future. Also, the high prevalence of type B cats demonstrates the possible risk of neonatal isoerythrolysis among stray cats living in İzmir, Turkey. © 2021, The Author(s).

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