Development of a safety protocol for training and using SARS-CoV-2 detection dogs: A pilot study.

Medical detection dogs have potential to be used to screen asymptomatic patients in crowded areas at risk of epidemics such as the SARS-CoV-2 pandemic. However, the fact that SARS-CoV-2 detection dogs are in direct contact with infected people or materials raises important concerns due to the zoonotic potential of the virus. No study has yet recommended a safety protocol to ensure the health of SARS- CoV-2 detection dogs during training and working in public areas. This study sought to identify suitable decontamination methods to obtain nonpathogenic face mask samples while working with SARS-CoV-2 detection dogs and to investigate whether dogs were able to adapt themselves to other decontamination procedures once they were trained for a specific odor. The present study was designed as a four-phase study: (a) Method development, (b) Testing of decon- tamination methods, (c) Testing of training methodology, and (d) Real life scenario. Surgical face masks were used as scent samples. In total, 3 dogs were trained. The practical use of 3 different decontam- ination procedures (storage, heating, and UV-C light) while training SARS-CoV-2 detection dogs were tested. The dog trained for the task alerted to the samples inactivated by the storage method with a sensitivity of100 % and specificity of 98.28 %. In the last phase of this study, one dog of 2 dogs trained, alerted to the samples inactivated by the UV-C light with a sensitivity of 91.30% and specificity of 97.16% while the other dog detected the sample with a sensitivity of 96.00% and specificity of 97.65 %.

Isolation and characterisation of Brucella melitensis by bacteriological and molecular methods from livestock in North Cyprus.

In this study, the isolation, biotyping and molecular characterisation of Brucella melitensis from cattle, sheep and goats in North Cyprus are reported on. A total of 319 raw milk samples obtained from seropositive dairy livestock (190 cattle, 74 sheep and 55 goats) and tissue samples including the liver, spleen and abomasal contents obtained from 32 aborted foetal samples (5 cattle, 18 sheep and 9 goats) were analysed for the presence and characterisation of the agent. B. melitensis was isolated and identified from 90 out of 319 (28.2%) milk and 19 out of 32 (59.4%) foetal samples by conventional bacteriological methods. Identification of all 109 isolates was confirmed by using real-time PCR with genus and species-specific primers. Following the preliminary identification, 27 selected isolates representing various counties and herds were further analysed by conventional methods. Twenty (74.1%) isolates were identified as B. melitensis biovar 1 and seven (25.9%) were identified as B. melitensis biovar 3. The Bruce-ladder multiplex PCR assay revealed that all the isolates were field strains. The results of the present study confirmed the presence of B. melitensis in livestock including the cattle population in North Cyprus. Even though the majority of the samples came from seropositive cattle, Brucella abortus was not isolated in the study. The results also revealed the potential public health risk of brucellosis in livestock emphasising the need of implementing strict control and eradication strategies against the disease in animal populations in order to protect human health.

Evaluation of PCR methods for detection of Brucella strains from culture and tissues.

The genus Brucella causes significant economic losses due to infertility, abortion, stillbirth or weak calves, and neonatal mortality in livestock. Brucellosis is still a zoonosis of public health importance worldwide. The study was aimed to optimize and evaluate PCR assays used for the diagnosis of Brucella infections. For this aim, several primers and PCR protocols were performed and compared with Brucella cultures and biological material inoculated with Brucella. In PCR assays, genus- or species-specific oligonucleotide primers derived from 16S rRNA sequences (F4/R2, Ba148/928, IS711, BruP6-P7) and OMPs (JPF/JPR, 31ter/sd) of Brucella were used. All primers except for BruP6-P7 detected the DNA from reference Brucella strains and field isolates. In spiked blood, milk, and semen samples, F4-R2 primer-oriented PCR assays detected minimal numbers of Brucella. In spiked serum and fetal stomach content, Ba148/928 primer-oriented PCR assays detected minimal numbers of Brucella. Field samples collected from sheep and cattle were examined by bacteriological methods and optimized PCR assays. Overall, sensitivity of PCR assays was found superior to conventional bacteriological isolation. Brucella DNA was detected in 35.1, 1.1, 24.8, 5.0, and 8.0% of aborted fetus, blood, milk, semen, and serum samples by PCR assays, respectively. In conclusion, PCR assay in optimized conditions was found to be valuable in sensitive and specific detection of Brucella infections of animals.

Investigation of aminoglycoside modifying enzyme genes in methicillin-resistant staphylococci.

Methicillin-resistant staphylococci may also be resistant to some other antibiotics as well as beta-lactams. In this study, co-existence of resistance to methicillin and aminoglycosides was genetically investigated in staphylococci. A total of 50 staphylococci from in-patients, 17 Staphylococcus aureus and 33 coagulase negative staphylococci (CNS) that contained mecA (gene encoding PBP 2a, an altered penicillin-binding protein) determined by polymerase chain reaction (PCR) were included in the study. Aminoglycoside modifying enzyme (AME) genes were investigated using multiplex-PCR. Aminocyclitol-6'-acetyltransferase-aminocyclitol-2''-phosphotransferase [aac(6')/aph(2'')] gene (encoding bifunctional acetyltransferases/phosphotransferases) was determined in 66% of the isolates, aminocyclitol-4'-adenylytransferase (ant(4')-Ia) gene (encoding phosphotransferases) in 24%, and aminocyclitol-3'-phosphotransferase (aph(3')-IIIa) gene (encoding nucleotidyltransferases) in 8%. Two isolates contained all these three genes. Thirty-six (72%) isolates had at least one of these genes. Three CNS and one S. aureus isolates sensitive to oxacillin had the mecA gene. In conclusion, a high rate of aminoglycoside resistance was determined in methicillin-resistant staphylococci. The aac(6')/aph(2'') was the most frequently detected.

Investigation of erythromycin and tetracycline resistance genes in methicillin-resistant staphylococci.

In this study, erythromycin [erm(A) and erm(C)] and tetracycline [tet(K) and tet(M)] resistance genes were investigated by multiplex polymerase chain reaction (PCR) in a total of 56 methicillin-resistant (mecA+) staphylococcal hospital isolates, 28 of which were determined to be Staphylococcus aureus (MRSA) and the other 28 were coagulase-negative staphylococci (MRCNS). Internal control primers amplifying a specific fragment of 16S rDNA of staphylococci were included in the multiplex PCR protocol to ensure the efficacy of amplification and to determine any PCR inhibition. No resistance genes were detected in 5 of 56 (8.9%) isolates in the study. In the study, tet(K) genes were detected widely (42.9%) in MRCNS, whilst tet(M) genes were detected in MRSA (50.0%). Regarding the erythromycin resistance genes, whilst erm(A) genes were detected in most (71.4%) MRSA isolates, detection rates of erm(C) genes were the same (64.3%) both in MRCNS and MRSA. The resistance rates for tetracycline and erythromycin were 57.1% and 78.6%, respectively, in MRSA isolates. In conclusion, in this study, the multiplex PCR technique including an internal control is shown to be a fast, sensitive, reliable, practical, reproducible and economic technique for the detection of erythromycin and tetracycline resistance in staphylococcal isolates.

[Investigation of methicillin resistance and aminoglycoside modifying enzyme genes in hospital staphylococci by multiplex-polymerase chain reaction]. / Hastane izolati stafilokoklarda metisiliN diRenç ve aminoglikozid modifiye edici enzim genlerinin multipleks-polimeraz zincir reaksiyonu ile aratirilmasi.

This study was conducted to investigate the presence of methicillin and aminoglycoside resistance encoding genes by multiplex-polymerase chain reaction (PCR) and by phenotypic methods in staphylococci isolated from inpatients' clinical specimens. The presence of aac(6')1aph(2"), aph(3')-IIIa and ant(4)-Ia genes encoding aminoglycoside modifying enzymes (AME) and mecA gene encoding methicillin resistance were genotypically investigated. A total of 19 S. aureus and 30 coagulase negative staphylococci (CNS) were tested. Thirty four (69.4%) of the isolates were found to be resistant to oxacillin with disk diffusion test, 33 (97%) of which were found to harbour mecA gene. The correspondance between oxacillin resistance and presence of mecA gene was found to be 100% in S. aureus isolates, while it was 95.7% in CNS. Twenty two (44.9%), 7 (14.3%) and 2 (4.1%) isolates were found to harbour aac(6')/aph(2"), aph(3')-IIIa and ant(4)-/a AME genes, respectively. At least one or more AME genes were detected in 72.7% of mecA positive isolates.

A case of aspergillosis in a broiler breeder flock.

A case of aspergillosis in a broiler breeder flock having respiratory and nervous system problems caused by Aspergillus fumigatus and Aspergillus niger is documented. Dyspnea, hyperpnea, blindness, torticollis, lack of equilibrium, and stunting were observed clinically. On postmortem examination of the affected birds, white to yellow caseous nodules were observed on lungs, thoracic air sacs, eyes, and cerebellum. Histopathologic examination of lungs and cerebellum revealed classic granulomatous inflammation and cerebellar lesions, necrotic meningoencephalitis, respectively. No lesions were noted in the cerebrum histopathologically. Aspergillus hyphae were observed in stained sections prepared from lesioned organs. Fungal spores and branched septate hyphae were observed in direct microscopy. Aspergillus fumigatus and A. niger were isolated from the inoculations prepared from the suspensions of organs showing lesions.