Detection of capnocytophaga canimorsus and capnocytophaga cynodegmi in dogs by cultural and molecular methods

Her yıl dünya çapında milyonlarca insanın hayvanlar tarafından ısırıldığı rapor edilmektedir. Isırıkların %90'ı köpekler tarafından meydana getirilir ve bu ısırık yaralarının büyük çoğunluğuna tıbbi müdahale yapılmamaktadır. Capnocytophaga cinsi içerisinde bulunan Capnocytophaga canimorsus ve Capnocytophaga cynodegmi türleri, köpeklerin ağız boşluğunda komensal olarak yaşayan mikroorganizmalardır ve bazen insanlarda ölümcül sistemik infeksiyonlara neden olabilmektedir. Bu tez çalışması ile köpeklerde oral florada görülen ve klinik tanısı rutin olarak konulamayan Capnocytophaga infeksiyonlarının tanıları CaL2, AS1, CaR ve CyR geni nükleotid sekansları kullanılarak yapılmıştır. Araştırmamızda Muğla ili ve çevresinde bulunan, diş taşı olan sahipli köpeklerden toplanan 200 adet oral svap örneği incelenmiş ve yapılan birinci PCR sonucunda 200 adet örneğin 11 (% 5.5)'inde CaL2 ve AS1 gen taşıyıcılığı saptanmıştır. İncelenen toplam 11 adet Capnocytophaga spp. pozitif örneğin 2 (% 18.2)'sinin sadece CaR geni açısından pozitif olduğu, 9 (% 81.8)'unun ise hem CaR, hem de CyR geni açısından pozitif olduğu tespit edilmiştir. Araştırma sonucunda, sadece CyR geni açısından pozitif olan örneğe ise rastlanmamıştır. Sadece CaR geni saptanan örnekler Capnocytophaga canimorsus pozitif, hem CaR, hem de CyR geni saptanan örnekler ise Capnocytophaga canimorsus ve Capnocytophaga cynodegmi pozitif olarak değerlendirilmiştir. Capnocytophaga canimorsus ve Capnocytophaga cynodegmi pozitif örneklerin 2 (% 22.2) adedi erkek hayvanlardan, 7 (% 78.2) adedi ise dişi hayvanlardan saptanmıştır. Sadece Capnocytophaga canimorsus pozitif olan 2 adet örneğin tamamı ise (% 100) dişi hayvanlardan saptanmıştır. Elde edilen pozitiflik oranlarının yaşa göre dağılımı incelendiğinde Capnocytophaga spp. pozitif olarak saptanan 11 örneğin 10 (% 90.9) adedinin 0-5 yaş grubu hayvanlardan, 1 (% 9.1) adedinin ise 6 yaşlı bir hayvandan saptandığı belirlenmiştir. Capnocytophaga canimorsus ve Capnocytophaga cynodegmi pozitif örneklerin 8 (% 88.8) adedi 0-5 yaş grubu hayvanlardan, 1 (% 11.2) adedi ise 6 yaşlı bir hayvandan saptandığı belirlenmiştir. Sadece Capnocytophaga canimorsus pozitif olan 2 adet örneğin tamamı ise 0-5 yaş grubu hayvanlardan saptanmıştır.Every year, it is reported that millions of people are bitten by animals worldwide. 90 % of the bites are caused by dogs and there is no any special medical care for these bite wounds. Capnocytophaga canimorsus and Capnocytophaga cynodegmi species are microorganisms found in Capnocytophaga genus and grow commensally in oral flora of dogs. Capnocytophaga canimorsus may also cause mortal systemic infections in human By this thesis work, the diagnosis of Capnocytophaga infections which could not made as routinely, was done by using CaL2, AS, CaR and CyR gene sequences. In our study, 200 oral swap samples which were taken from owned dogs which have dental plaque, were examined and in the end of first PCR, CaL2 and AS1 gene were detected from 11 (5.5 %) samples out of 200 samples. It is found that 2 (18.2 %)of the samples were positive for CaR gene and 9 (81.8 %) of the samples were positive for both CaR and CyR gene out of examined 11 Capnocytophaga spp. positive samples. As a result of this research, there was no any positive sample detected that is only positive for CyR gene. The samples that only CaR positive were evaluated as Capnocytophaga canimorsus positive, the samples that both CaR and CyR positive were evaluated as Capnocytophaga canimorsus and Capnocytophaga cynodegmi positive. 2 (22.2 %) of the Capnocytophaga canimorsus and Capnocytophaga cynodegmi positive samples were detected from male animals, 7 (78.2 %) of them were detected from female animals. The whole 2 (100 %)samples that are only Capnocytophaga canimorsus positive were detected from female animals. In the result of positivity range by age, 10 (90.9 %) of the samples were detected from 0-5 age animals, and 1 (9.1 %) of the samples were detected from 6 aged animal out of 11 Capnocytophaga spp. positive samples. 8 (88.8 %) of the samples were detected from 0-5 age animals, and 1 (11.2 %) of the sample were detected from 6 aged animal out of 9 Capnocytophaga canimorsus and Capnocytophaga cynodegmi positive samples. The whole 2 (100 %) samples that are only Capnocytophaga canimorsus positive were detected from 0-5 age animals

Kısırlaştırılmış Pitbull ırkı köpeklerin oral florasında periodontal patojenlerin varlığının araştırılması

Dental plaque bacteria are responsible for the development of periodontal disease in dogs. The aim of this study was to determine the presence of pathogenic bacteria associated with periodontal disease in the dental plaques of spayed Pitbull dogs. Plaque and subgingival swap samples were obtained from 27 male and 31 female dogs. Samples were analyzed with PCR for Porphyromonas gingivalis, Porphyromonas gulae, Treponema denticola, Tannerella forsythia, Capnocytophaga ochracea, Capnocytophaga sputigena, Prevotella intermedia, Prevotella nigrescens, Campylobacter rectus, Aggregatibacter actinomycetemcomitans, and Eikenella corrodens. Positive samples (52/58) with 16S rRNA PCR were determined as 89.65% and 6 samples were negative. All samples were confirmed except P.nigrescens and A.actinomycetemcomitans. P.gingivalis (50/52) was detected in 96.15% and P.gulae and P.intermedia (33/52) were in 63.46%. 1.9% of T. denticola and C. rectus (1/52) were confirmed only in male dogs. Also, gender did not affect the presence of pathogenic bacteria. As a result of the research, bacterial variations strongly associated with Pitbull periodontal diseases were obtained.</p

Keratokonjunktivitisli Kedilerden Bakteriyel Patojenlerin İdentifikasyonu ve Antimikrobiyal Duyarlılıklarının Belirlenmesi

Keratokonjunktivitis görülen kedilerde bakteriyelpatojenler sıklıkla görülmektedir. Bu çalışmanın amacı pürülan inflamasyon ve konjunktivalhiperemi gösteren 20 hasta kedi ile semptomsuz 5 sağlıklı kedinin konjunktivalflorasından bakteriyel patojenlerin identifikasyonunu gerçekleştirmek veizolatların antimikrobiyal duyarlılıklarını belirlemektir. Kedilerin konjunktivalkesesinden svap örnekleri alındı ve %5 kanlı agar üzerine inoküle edildi.Aerobik ve mikroaerofilik (%7 CO2) ortamda 24-48 saat 37°C'de inkübeedildi. Konjunktivitisli kedi svapörneklerinden en sık Koagulaz NegatifStaphylococcus (%22,5), Moraxella sp. (%20) ve Streptococcus sp. (%17,5) türleri, sağlıklıkedi svap örneklerinden ise Klebsiellasp. ve Bacillus sp. (%23) türleriyüksek oranda izole edilmiştir. İzolatlarda gentamisin, basitrasin, amoksisilinklavulanik asit (%75) ile siprofloksasin'e (%65) duyarlılık, streptomisin (%48),penicillin G (%53) ve tetrasiklin'e (%42) ise dirençlilik tespit edildi. Kedilerinkeratokonjunktivitis infeksiyonlarında mikrobiyal patojen çeşitliliğinintanı'da dikkate alınması ve hastalık tedavisi için antimikrobiyallerin akıllıcaseçilmesi faydalı olacaktır.</p

İnfeksiyöz Deve (Camelus dromedarius) Keratokonjonktivitis Olgularında Moraxella bovoculi'nin İdentifikasyonu ve Antimikrobiyal Duyarlılıklarının Belirlenmesi

İnfeksiyöz keratokonjonktivit (İKC) sığır ve koyunlarda ekonomik kayıplara neden olan birgöz hastalığıdır. Hastalık etkenleri genel olarak Moraxella bovis ve M. ovis olarak bilinirken2007'de İKC'den sorumlu türler arasına M. bovoculi ’de tanımlanmıştır. Bu çalışmanın amacıkonjonktival hiperemi, oküler ağrı, fotofobi ve lakrimasyon semptomları gösteren develerdeMoraxella spp. varlığının saptanması ve antimikrobiyal duyarlılık profillerindeki farklılıklarıbelirlemektir. Aydın yöresinde 30 adet devenin (Camelus dromedarius) sağ ve sol gözlerindenbilateral (n= 60 örnek) konjonktival svap örnekleri toplandı. Sürüntü örneklerinin %10(6/60)’nundan Moraxella spp. (6/60; %10) izole edildi. Biyokimyasal olarak nitrat redüksiyonpozitif olanlar M. ovis ve M. bovoculi (M. bovis negatif) ile uyumluluk gösterdi. Ayrıca, 357F ve1492R evrensel primerleri kullanarak 16S rRNA PCR gerçekleştirildi ve nükleotit sekansıyapıldı. Sanger sekanslama ile izolatların Moraxella bovoculi (Moraxella bovoculi suşu 3709'a%98-99 benzerlik Access. No: GU181221.1) olduğu doğrulandı. İzolatlarda eritromisin (%100),amoksisilin-klavulanik asit, penisilin, siprofloksasin ve tetrasiklin (%67) gibi yaygınantibiyotiklere direnç, sefotaksim, gentamisin ve imipeneme (%100) duyarlılık tespit edildi. M.bovoculi suşu develerin göz enfeksiyonlarında ülkemizde daha önce rapor edilmemiştir. Bunedenle çalışmamız develerin göz enfeksiyonlarında M. bovoculi'nin varlığını doğrulamaktadırve develerin göz enfeksiyonlarından izole edilebileceğine vurgu yapmaktadır.&nbsp;: Infectious keratoconjunctivitis (IKC) is an eye disease that causes economic losses incattle and sheep. The causative agents of disease are commonly known as Moraxella bovis andM. ovis. In 2007, M. bovoculi was also identified among the species responsible for IKC. The aimof this study was to detect Moraxella spp. in camels with conjunctival hyperemia, ocular pain,photophobia, and lacrimation symptoms. The aim of this study is to detect the presence ofantimicrobial susceptibility profiles and to determine the differences in antimicrobialsusceptibility profiles. Bilateral (n= 60 samples) conjunctival swab samples were collected fromthe right and left eyes of 30 camels (Camelus dromedarius) in the Aydin region. Moraxella spp.(6/60; 10%) strains were isolated from swab samples by phenotypic and genotypic methods.Biochemically nitrate reduction and gelatinase positive M. ovis and M. bovoculi (M. bovisnegative) showed compatibility. Also, 16S rRNA PCR was performed using 357F and 1492Runiversal primers, and nucleotide sequencing was performed. The isolates were confirmed to beM. bovoculi (98-99% similarity to M. bovoculi strain 3709 Access. No: GU181221.1) by Sangersequencing. Resistance to erythromycin (100%), amoxicillin-clavulanic acid, penicillin,ciprofloxacin, and tetracycline (67%) and susceptibility to cefotaxime, gentamicin, and imipenem(100%) were detected in the isolates. M. bovoculi strain has not been previously reported in camel&nbsp;eye infections in our country. Therefore, our study confirms the presence of M. bovoculi in cameleye infections and emphasizes that camels can be isolated from eye infections&nbsp;</p

Improving Recombinant Membrane Protein Expression in Rhodobacter sp. via Utilization of MISTIC Protein Fusion

Being located at the interface of intracellular and extracellular regions of biological cells, membrane proteins (MPs) mediate many vital events including signal transduction, energy generation, selective transport of solutes etc.. Because of these regulatory properties, nearly 70% of currently FDA approved drugs on the market target membrane proteins. Despite their importance, low abundance of MPs in natural resources is one of the major bottlenecks in studying their structure and functionality. Moreover, MPs have been employed in miscellaneous applications such as sensor technology, optogenetics, biomimetic separation membranes, in-vitro photosynthesis leads to an ever increasing demand for avaliability of MPs. Recombinant expression of MPs from heterologous expresion hosts was proven to be challenging especially for mammalian MPs due to improper folding, inclusion body formation and toxic effects on host organism. Therefore, development of alternative expression host systems for functional MP expression is of critical importance. Purple non-sulphur bacteria from Rhodobacter genus are known for their diverse metabolic pathways and intensively studied as model organisms in photosynthetis research. Under anaerobic, photosynthetic growth mode, plasma membrane surface area of Rhodobacter ssp immensely increase to accomodate the photosynthetic apparatus. An order of magnitude higher membrane surface area with inducible expression vectors imparts Rhodobacter sp as an attractive recombinant MP expression host. In this study, we employed codon optimized MISTIC chaperonin protein gene as N-terminal and mBanana fluorescent protein gene as C-terminal fusion partner for human AQP6 and AQP9 genes for co-expression in Rhodobacter sp. Cloning procedures were carried out in E. coli Top10 and expression vectors were transfered in Rhodobacter by biparental mating using E. coli S17 -pir. Protein expression was investigated under semi-aerobic and photosynthetic growth modes using two different promoters. A significant improvement in expression titers was observed with MISTIC fusions as monitored by SDS-PAGE.</p

Isolation, Characterization and Antimicrobial Activity of Bacteriocin Producing Lactic Acid Bacteria from Raw Water Buffalo Milk

Antibiotic resistance is a globally emerging health care concern. In recent years, a major difficulty in treatment of diseases such as tuberculosis, malaria, pneumonia and certain nosocomial infections is the presence of multi drug-resistant pathogens. In this regard, availability of the alternative antimicrobial materials is of paramount importance. Bacteriocins are ribosome-derived antimicrobial peptides generated by vast majority of bacteria to gain advantage over their competition. Bacteriocins from lactic acid bacteria (LAB) are often employed as natural food preservatives in food industry to increase the shelf-life of products. Their utilization as antimicrobial and anti-cancer agents in medical and veterinary treatments have been proposed. Therefore, discovery of new bacteriocin-producer LAB strains will benefit health and food industry applications. In this study, bacteriocin producer LAB strains were isolated from water buffalo milk samples collected from dairy farms located in Sorgun and Akdağmağdeni districts of Yozgat Province. Selective culture media were used for isolation of LAB genera including Lactobacillus, Enterococcus and Bifidobacterium. Screening of bacteriocin producer strains were carried out by reverse-side agar spot test using E. coli and S. aureus as indicator bacteria. A further verification of bacteriocin activity was carried out using neutralized cell-free supernatants via agar well diffusion assay. LAB isolates were characterized using miscellaneous biochemical methods including TSI, MR-VP, indole and catalase tests. Majority of the bacteriocin producer strains formed some degree of inhibition zone against selected indicator bacteria implying a broad bacteriocin antimicrobial activity spectrum, which might enable their utilization as novel antimicrobial agents.</p

Otolog Trombosit Konsantrelerinin İmmunolojik ve Antimikrobiyal Etkileri

Antibiotic resistance has remarkable potential in human beings and veterinary medicine. However, to prevent the clinical reflection of this resistance from reaching the feared dimensions, there is a requirement for antimicrobial treatment options supported and improved with new molecular biocursors at the preclinical point. Platelet-rich plasma (PRP) and fibrin (PRF) are biomaterial products that recently used to increase the anti-infective defense system by platelet growth factors to support postoperative wound healing, bone regeneration, graft stabilization, biofilm inhibition, catheter hygiene, and hemostasis. Recently, research has been carried out on antibacterial, antifungal, and prevention of clinical biofilm formation. Autologous platelet concentrates are autogenous and do not cause any immunological reaction or infection. Therefore, the choice and application of regenerative therapies are being favored due to their nominal invasive procedures. In particular, PRP and PRF are of interest because of their influence to stimulate and speed up the injury area healing process. Cytokines and growth factors involved in the formation of PRP are played an important role in the recovery process. This article aims to evaluate the antibacterial, antifungal and antibiofilm properties of PRP and PRF in the field of microbiology. In addition, the act of growth factors in the process of healing and their use in regenerative treatments were also evaluated.&nbsp;</p

Ege Bölgesinde Koyun ve Kuzu Pnömonisi Olgularında Pasteurella multocida ve Mannheimia haemolytica İzolasyonu ve Antibiyotik Dirençlerinin Tespiti

In this study, the presence and antimicrobial susceptibility of Mannheimia haemolytica and Pasteurella multocida pathogens in 200 sheep-lamb lung samples with pneumonia were investigated in 7 provinces of the Aegean region between 2019-2021. The tissues of the animals were inoculated into 7% blood agar and then incubated at 37oC for 24-48 hours. Gram staining was performed on the pure colonies and Gram-negative, oxidase-positive samples were confirmed with the Vitek 2 system. An Antibiogram test was performed and amoxicillin-clavulanic acid, enrofloxacin, erythromycin, florfenicol, gentamicin, oxytetracycline, sulfamethoxazole-trimethoprim, tulathromycin discs were used. M. haemolytica was detected in 20(10%) of the samples and P. multocida was detected in 22(11%) of the samples. 20 of the P. multocida isolates (91%) were susceptible to amoxicillin-clavulanic acid, florfenicol, and tulathromycin, and 20 of the M. haemolytica isolates (100%) were susceptible to enrofloxacin, oxytetracycline, florfenicol, and tulathromycin. Subsequently, 6 of the P. multocida isolates (27%) were resistant to erythromycin and oxytetracycline, and 5 of the M. haemolytica isolates (25%) were to sulfamethoxazole-trimethoprim. As a result, M. haemolytica and P. multocida can be seen with similar percentages as pneumonic agents in sheep-lamb pneumonia cases in the Aegean region, but effective treatment can be done with the right antibiotic selection.</p

Ege bölgesi neonatal kuzu ölümlerinde Escherichia coli septisemisinin ve antibiyotik duyarlılığının araştırılması

Bu çalışmada 2019-2021 yılları arasında Ege bölgesine ait 7 ilde (Aydın, Denizli, İzmir, Kütahya, Manisa, Muğla, Uşak) görülen neonatal (0-28 gün) kuzu ölümlerinde Escherichia coli (E. coli) septisemisi araştırıldı ve etkenin antimikrobiyal duyarlılıkları tespit edildi. Araştırmanın materyalini 150 adet kuzu viseral organ ve dokusu (akciğer, karaciğer, dalak, lenf, kemik iliği ve barsak) oluşturdu. Örnekler, nutrient broth 37°C’de 24 saat aerobik şartlarda inkube edildi ve daha sonra %7 kanlı agara ve MacConkey agara ekimler yapılarak 37oC'de 24-48 saat inkubasyona bırakıldı. Kanlı agarda grimsi S tipli, MacConkey agarda pembe, mukoid olmayan koloni oluşturan Gram negatif basiller E. coli olarak değerlendirildi ve Vitek 2 sistemi ile de doğrulandı. Örneklerin %88,66 (133/150)'sında E. coli etkeni tespit edildi. Etken izolasyonu en çok İzmir (31/133; %23,30) ve Aydın (25/133; %18,80)'da yapıldı. Diğer illerde bulgular birbirine yakın seyir gösterdi. Antibiyotik duyarlılık testinde amoksisilin- klavulanik asit (30 µg), sefoperazon (30 µg), eritromisin (15 µg), penisilin G (10 units), gentamisin (10 µg), tetrasiklin (30 µg), trimetoprim-sulfametoksazol (25 µg) ve enrofloksasin (5 µg) ticari diskleri kullanıldı. İzolatların 110’u (%73,33) gentamisin'e, 80'i sefoperazon'a (%53,33) ve 70'i (%46,66) amoksisilin-klavulanik asit'e duyarlı bulundu. Ek olarak, izolatların tüm'ü penisilin G' ye (%100), 146'sı (%97,33) eritromisin'e, 122'si tetrasiklin'e (%81,33) ve 119'u (%79,33) sulfametaksazol-trimethoprim'e dirençli bulundu. Sonuç olarak Ege bölgesinde iç organlar tutulumu ile karakterize E. coli septisemisi görülmektedir. Mortalite'nin antibiyogram ile akılcı antibiyotik kullanımı, doğru tedavi yaklaşımları ve koruyucu hekimlik uygulamaları ile azalma göstereceği ve ekonomiye olumlu katkı sağlayacağı düşünülmektedir.In this study, Escherichia coli (E. coli) septicemia was investigated in neonatal (0-28 days) lamb deaths in 7 provinces of the Aegean region (Aydın, Denizli, İzmir, Kütahya, Manisa, Muğla, Uşak) between 2019-2021, and antimicrobial susceptibility was determined. The material of the study consisted of 150 lamb visceral organs and tissues (lung, liver, spleen, lymph, bone marrow and intestine). The samples were incubated in nutrient broth under aerobic conditions and then inoculated on 7% blood agar and Macconkey agar, and incubated at 37°C for 24-48 hours. Gram-negative bacilli that form grayish S-type on blood agar and pink, non-mucoid colony on Macconkey agar were evaluated as E. coli and were also confirmed by the Vitek 2 system. E. coli agent was detected in 88.66% (133/150) of the samples. Agent isolation was most common in İzmir (31/133; 23.30%) and Aydın (25/133; 18.80%). Findings in other provinces showed a similar trend. In antibiotic susceptibility test, amoxicillin-clavulanic acid, cefoperazone, erythromycin, penicillin G, gentamicin, tetracycline, trimethoprim-sulfomethoxazole and enrofloxacin commercial discs were used. Of the isolates, 110 (73.33%) were sensitive to gentamicin, 80 (53.33%) to cefoperazone, and 70 (46.66%) to amoxicillin-clavulanic acid. In addition, all of the isolates were resistant to penicillin G (100%), erythromycin (97.33%), tetracycline (81.33%), and sulfamethoxazole-trimethoprim (79.33%). As a result, E. coli septicemia characterized by visceral involvement is seen in the Aegean region. It is thought that mortality will decrease with the right treatment approaches and rational antibiotic use and will contribute positively to the economy.</p

Presence of zoonotic black-pigmentedperiodontal pathogens in the oral microbiotaof pet and stray cats

Black-pigmented bacteria are one of&nbsp;the neglected species to&nbsp;cause periodontal disease in&nbsp;cats, and theyare also zoonotic agents that pose an&nbsp;infection risk to&nbsp;humans. In&nbsp;this study, we&nbsp;aimed to&nbsp;determine the&nbsp;presenceof&nbsp;Porphyromonas gingivalis, Porphyromonas gulae and Prevotella nigrescens in&nbsp;the oral microbiota of&nbsp;pet and straycats. Dental swab samples were taken from 25&nbsp;pet cats and 25&nbsp;stray cats with symptoms of&nbsp;periodontal disease andthen investigated by&nbsp;multiplex polymerase chain reaction using 16S&nbsp;rRNA species-specific primers. As&nbsp;a&nbsp;result of&nbsp;themultiplex PCR analysis, P.&nbsp;gingivalis 3/25 (12%), P.&nbsp;nigrescens 1/25 (4%), P.&nbsp;gingivalis&nbsp;+ P.&nbsp;gulae&nbsp;7/25 (28%), P.&nbsp;gingivalis&nbsp;+ P.&nbsp;nigrescens 1/25 (4%), P.&nbsp;gulae&nbsp;+ P.&nbsp;nigrescens 1/25 (4%), and P.&nbsp;gingivalis&nbsp;+ P.&nbsp;gulae&nbsp;+ P.&nbsp;nigrescens&nbsp;2/25(8%) were molecularly typed in&nbsp;the pet cats. In&nbsp;addition, 1/25 (4%) of&nbsp;P.&nbsp;gulae and 21/25 (84%) of&nbsp;P.&nbsp;gingivalis&nbsp;+P.&nbsp;gulae were typed in&nbsp;the stray cats. In&nbsp;10/25 (40%) pet and 3/25 (12%) stray cat samples, no&nbsp;bacteria were detectedby&nbsp;molecular typing. In&nbsp;summary, the results provide strong evidence that black-pigmented zoonotic pathogensare associated with cat periodontal disease.</p