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Fulltext Improving the methods for isolation of monocyte and establishing macrophage cell culture in caprine model

Shaqinah NN, Mazlina M, Zamri-Saad M, Hazilawati H, Jasni S

Cytotechnology, 2016 Aug;68(4):1655-9.
PMID: 25511802 DOI: 10.1007/s10616-014-9836-7

Monocytes are widely used for immunological research, especially in the study of innate immune system. Although methods for isolation of human monocytes have been established, the procedure for non-human monocyte has not been well developed. This paper describes an improved method for isolation of monocyte and the subsequent macrophage cultivation from caprine blood. Monocytes were isolated from 16 ml of heparinized caprine blood using double density methods; the Ficoll and Percoll. The number of monocytes obtained was 5.12 ± 0.89 × 10(7) cells/ml at 70 % purity. The isolated monocytes were maintained in 10 % fetal bovine serum-enriched Dulbecco's Modified Eagle Medium for maturation to form macrophage cell culture. At the end of the experiment, the harvested macrophage was 2.48 ± 0.33 × 10(6) cells/ml.
Fulltext Pathological changes and bacteriological assessments in the urinary tract of pregnant goats experimentally infected with Brucella melitensis

Mazlina M, Khairani-Bejo S, Hazilawati H, Tiagarahan T, Shaqinah NN, Zamri-Saad M

BMC Vet Res, 2018 Jun 25;14(1):203.
PMID: 29940976 DOI: 10.1186/s12917-018-1533-x

BACKGROUND: This study was conducted to investigate the pathological changes and distribution of B. melitensis in the urinary tract of pregnant goats following acute experimental infection. Six Jamnapari crossbred does in their third trimester of pregnancy were randomly assigned into two groups; Group 1 was uninfected control and Group 2 was inoculated conjunctival with 0.1 mL of the inoculums containing 109 cfu/mL of live B. melitensis. All does were sacrificed 30 days post-inoculation before the kidney, ureter, urinary bladder, urethra and vaginal swab were collected for isolation of B. melitensis. The same tissue samples were fixed in 10% neutral buffered formalin for hematoxylin and eosin, and immunoperoxidase staining.
RESULTS: None of the goats showed clinical signs or gross lesions. The most consistent histopathology finding was the infiltration of mononuclear cells, chiefly the macrophages with few lymphocytes and occasionally neutrophils in all organs along the urinary tract of the infected goats of Group 2. Other histopathology findings included mild necrosis of the epithelial cells of the renal tubules, congestion and occasional haemorrhages in the various tissues. Kidneys showed the most severe lesions. Immunoperoxidase staining revealed the presence of B. melitensis within the infiltrating macrophages and the epithelium of renal tubules, ureter, urethra and urinary bladder. Most extensive distribution was observed in the urinary bladder. Brucella melitensis was successfully isolated at low concentration (3.4 × 103 cfu/g) in the various organs of the urinary tract and at high concentration (2.4 × 108 cfu/mL) in the vaginal swabs of all infected goats. Although B. melitensis was successfully isolated from the various organs of the urinary tract, it was not isolated from the urine samples that were collected from the urinary bladder at necropsy.
CONCLUSION: This study demonstrates the presence of low concentrations of B. melitensis in the organs of urinary tract of pregnant does, resulting in mild histopathology lesions. However, B. melitensis was not isolated from the urine that was collected from the urinary bladder.
Streptococcus agalactiae isolates from cultured fishes in Malaysia manifesting low resistance pattern towards selected antibiotics

Aisyhah MA, Amal MN, Zamri-Saad M, Siti-Zahrah A, Shaqinah NN

J Fish Dis, 2015 Dec;38(12):1093-8.
PMID: 25704397 DOI: 10.1111/jfd.12351
Antigenic distribution, pathological changes, antibody response and serological detection in non-pregnant goats following experimental infection by Brucella melitensis

Mazlina M, Khairani-Bejo S, Hazilawati H, Shaqinah NN, Zamri-Saad M

Transbound Emerg Dis, 2021 Jul;68(4):2028-2038.
PMID: 32979887 DOI: 10.1111/tbed.13850

This study describes the pathological changes, antibody response, isolation and distribution patterns following exposure of non-pregnant goats to live Brucella melitensis. Eighteen healthy adult female goats were divided into two equal groups. Group 1 was infected via conjunctival sac with 109 cfu/ml of B. melitensis, while Group 2 was similarly exposed to sterile PBS. Serum and swabs from the eyes and vagina were collected at 5-day intervals. On days 15, 30 and 75 post-infection, 3 goats from each group were killed before the conjunctiva, ovary, oviduct, uterine horn, uterine body and vagina, the submandibular, prescapular and supramammary lymph nodes, the mammary gland, liver, spleen, urinary bladder and synovial membranes were collected for bacterial isolation and pathological study. Exposure of non-pregnant goats to B. melitensis did not produce clinical signs and gross lesions but produced mild necrosis and inflammation in the lymph nodes, the organs of reproductive tract, the mammary gland and urinary bladder. In general, microscopic lesions were most severe in the D75 goats, followed by D30 and D15 goats. Brucella melitensis was most frequent and significantly (p
The effects of fish gender on susceptibility to acute Streptococcus agalactiae infection in Javanese medaka Oryzias javanicus

Amal MNA, Zarif ST, Suhaiba MS, Aidil MRM, Shaqinah NN, Zamri-Saad M, et al.

Microb Pathog, 2018 01;114:251-254.
PMID: 29217326 DOI: 10.1016/j.micpath.2017.11.069

This study describes the susceptibility of different fish gender following acute Streptococcus agalactiae infection by using Javanese medaka Oryzias javanicus as test fish. The fish were grouped into four groups, which were: (1) all-male; (2) all-female; (3) mixed-gender (1 male: 1 female ratio); and (4) control non-infected (1 male: 1 female ratio). The fish in group 1, 2 and 3 were intraperitoneally exposed to 5.4 × 108 CFU/mL of S. agalactiae, while for group 4, the fish were exposed using sterile broth. The main clinical signs and histopathological changes of infected Javanese medaka were commonly observed in S. agalactiae infected fishes. However, no difference on clinical signs and histopathological changes of fish in group 1, 2 and 3 were noticed. The Javanese medaka mortality in group 1, 2 and 3 were observed from 4 h post infection (hpi) to 6 hpi, with the cumulative mortality from 3% to 30%. Then, the mortality increased at 12 hpi, with the range from 53% to 80%. However, 100% of the infected fish dead at 24 hpi. No clinical sign, histopathological change and fish mortality recorded in group 4. Generally, the clinical signs, mortality patterns, cumulative mortality and histopathological changes of Javanese medaka infected by S. agalactiae did not show any difference between the all-male, all-female and mixed-gender groups. This indicates that the susceptibility of fish to S. agalactiae infection is not influenced by their gender.