Detection of colistin-resistant Escherichia coli isolated from broiler chickens in Kelantan, Malaysia.

Antimicrobial resistant Escherichia coli have become an ever increasing problem in human, and animal health and production. The imprudent use of antibiotics and poor hygienic practices especially in poultry industries have been contributing to the emergence and spread of E. coli species resistant to broad spectrum antibiotics including Colistin. This study was conducted to detect colistin - resistance and antibiotic sensitivity patterns in E. coli isolated from broiler chickens in Kelantan. A total of 320 cloacal swabs were collected from apparently healthy broiler chickens in different districts of Kelantan and were analysed using routine microbiological methods, Kirby-Bauer method for antimicrobial susceptibility test and PCR amplification of species-specific and colistin - resistance encoding genes. Out of the 320 samples, 91 isolates were confirmed as E. coli and 21/91 (23.08%) were positive for colistin - resistant encoding gene, mcr-1. Most of the isolates were resistant to tetracycline (95.24%), chloramphenicol (85.71%), and sulphamethoxazole/ trimethoprim (85.71%). However, the isolates were less resistant towards piperacillin/ tazobactam (4.76%) and meropenem (9.52%). The findings from this study reveal the emerging threats of colistin - resistant in local food animal production, particularly in poultry production industry. However, more comprehensive, and large-scale studies focusing on more resistance patterns using determination of minimum inhibitory concentration (MIC), virulence and resistance characteristics and molecular epidemiology of colistin - resistant E. coli are recommended for better understanding of the epidemiology and to implement the appropriate control and prevention strategies.

Molecular investigation of Mycoplasma haemofelis in stray cats in Kota Bharu, Kelantan.

Mycoplasma haemofelis (M. haemofelis) is a gram-negative, epicellular bacteria that infects feline red cells (RBC). The pathogen appears as small blue cocci, rings, or rods on the edges or across the surface of RBCs and often causes haemolytic anemia. However, the epizootiology of M. haemofelis is still poorly understood. So far, there are only a few studies that have been carried out to determine the prevalence of M. haemofelis. Most of the studies were conducted in Europe and other continents and the epidemiology of M. haemofelis in Malaysia, particularly in stray cats have not been reported. In this study, 60 blood samples were collected from stray cats in Kota Bharu and were examined by using thin blood smear and polymerase chain reaction (PCR) methods. This study showed that seven out of the 60 blood samples were positive for M. haemofelis by using PCR. Thus the prevalence rate of M. haemofelis in stray cats in Kota Bharu from this study was 11.7%. However, more studies with larger sample size and diverse sample distribution should be conducted to better understand the occurrence of this pathogen in both housed and stray cats. Moreover, the genetic variability of M. haemofelis infecting domestic and wild animals need to be conducted to verify the relationship among geographic distribution, genetic diversity and the potential threats to animal and human health.

Molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) among veterinary students and personnel at a veterinary hospital in Malaysia.

In this study, we report the molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) among veterinary students and personnel in Malaysia. Nasal and oral swabs were collected from 103 veterinary medicine students and 28 personnel from a veterinary hospital. Antibiotic sensitivity test (AST), minimum inhibitory concentration (MIC) test, and PCR amplifications of nucA and mecA gene were performed. Molecular characterization of the isolates was conducted using multilocus sequence typing (MLST), staphylococcal protein A gene (spa) typing, and pulsed-field gel electrophoresis (PFGE). Results from MLST show the presence of the pandemic and widespread MRSA clones, ST5 and ST59. Spa gene typing revealed spa type t267 which has a wide geographical distribution. A new spa type, t5697 was found in this study. Fingerprint analysis by using PFGE show heterogeneity of the isolates. These findings affirm the importance of MRSA in veterinary settings and underscore the need for further extensive research to devise contextual control and prevention strategies.

Long-term survival in multiple myeloma is associated with a distinct immunological profile, which includes proliferative cytotoxic T-cell clones and a favourable Treg/Th17 balance.

Despite improved outcomes in multiple myeloma (MM), a cure remains elusive. However, even before the current therapeutic era, 5% of patients survived >10 years and we propose that immune factors contribute to this longer survival. We identified patients attending our clinic, who had survived >10 years (n=20) and analysed their blood for the presence of T-cell clones, T-regulatory cells (Tregs) and T helper 17 (Th17) cells. These results were compared with MM patients with shorter follow-up and age-matched healthy control donors. The frequency of cytotoxic T-cell clonal expansions in patients with <10 years follow-up (MM patients) was 54% (n=144), whereas it was 100% (n=19/19) in the long-survivors (LTS-MM). T-cell clones from MM patients proliferated poorly in vitro, whereas those from LTS-MM patients proliferated readily (median proliferations 6.1% and 61.5%, respectively (P<0.0001)). In addition, we found significantly higher Th17 cells and lower Tregs in the LTS-MM group when compared with the MM group. These results indicate that long-term survival in MM is associated with a distinct immunological profile, which is consistent with decreased immune suppression.

Phenotypic and genotypic characterization of methicillin-resistant Staphylococcus aureus (MRSA) isolated from dogs and cats at University Veterinary Hospital, Universiti Putra Malaysia.

Methicillin-resistant Staphylococcus aureus (MRSA) is known to cause nosocomial infections and is now becoming an emerging problem in veterinary medicine. The objective of the study was to determine the presence of MRSA in 100 cats and dogs sampled between November 2007 and April 2008 at the University Veterinary Hospital, Universiti Putra Malaysia. MRSA was detected in 8% of pets sampled. Ten percent (5/50) and 6% (3/50) of the isolates were from dogs and cats, respectively. All MRSA isolates possessed the mecA gene and were found to be resistant to at least three antimicrobials with a minimum of Oxacillin MIC of 8 µg/mL. One isolate (CT04) had an extremely high MIC of >256 µg/mL. The MLST type ST59 found in this study have been reported earlier from Singapore and other countries as a strain from animal and community-associated MRSA respectively. Pulsed-field gel electrophoresis revealed five pulsotypes. Two isolates from cats (CT27 and CT33) and three isolates from dogs (DG16, DG20, and DG49) were respectively assigned to pulsotypes B and D. The study suggests that cats and dogs in Malaysia are potential reservoirs for MRSA.

Characterisation and relevance of CD138-negative plasma cells in plasma cell myeloma.

INTRODUCTION: The use of CD138 to isolate CD138(+) plasma cells (PCs) from plasma cell myeloma (PCM) patients' bone marrow samples has been used extensively in myeloma research. We sought to highlight the problem with this selection process, by demonstrating that a subpopulation of CD138⁻ plasma cells exists which is not included in these analyses. METHODS: Retrospective analysis of a patient database was carried out on all PCM patient bone marrow biopsies taken between 4/9/07 and 18/2/09 (n = 218). CD138(+) and CD138⁻ cell populations were separated using flow cytometry cell sorter then analyzed for percentage of cells in S phase using plasma cell labeling index as an indicator of proliferation. RESULTS: Database results indicated a CD138⁻ PC population in all PCM patient samples which also had a significantly increased (r = 0.53; P < 0.0001) CD45 expression, an indicator or immaturity. Flow cytometric analysis demonstrated the presence of a more immature, higher proliferating CD138⁻ PC population through a significantly (t = 3.26; P < 0.02) higher number of CD138⁻ PCs in S phase compared with the CD138(+) cells. CONCLUSION: We have characterised the CD138⁻ PCs as more immature and with a significantly higher proliferative potential. The current trend to ignore this more immature and proliferative subpopulation of malignant PCs may have serious implications when determining gene expression, classifications and drug sensitivity of the malignancy.