Evaluation of an improved Giemsa staining technique for blood borne parasite in thick and thin blood film from dogs

Many a times the use of rapid diagnostic tests for blood borne parasites like trypanosomiasis and Babesiosis is increasingly being used, but the gold standard for its detection is still the use of microscopy both as reference and confirmatory diagnosis. To determine the effectiveness of the adjusted stock giemsa staining technique over the conventional methods. Venous Blood samples were collected from 10 dogs in EDTA and then used for the simultaneous preparation of two thin and thick smear slides, one stained according to Giemsa 1:20 dilution for 30 mins while the other was stained using the Stock Solution for 30seconds the diluted with buffered saline for 20seconds and rinsed. Fixation of the thin smear was done in a covered staining jar containing anhydrous methanol for 1 to 2 min, after which the slides were air-dried. From the result obtained from 10 dogs blood samples gotten from the veterinary clinic, the adjusted giemsa staining technique showed a positive differentiation when compared to the 1:20 dilution, a total of 7 blood samples tested positive for blood borne parasites, Trypanosoma evansi, Babesiosis cani and Heart worm. The highest percentage occurrence was T.evansi (40%), Babesiosis cani(20%) and Heart worm (10%).The adjusted Giemsa staining technique serves as a fast, easy and less complex alternative to the 1:20 dilution, where the solution has to be diluted from the stock solution and then stained, although the only disadvantage to this technique would be easy contamination of the stock solution, but the advantages here is that it saves time, quicker result output and better differentiation microscopically.

Canine trypanosomosis: a case report

Trypanosomosis is a haemoprotozoan infection affecting a broad range of wild and domestic animals including dogs. Infected dogs may die within 2-4 weeks in the acute and fatal form of the disease. A dog was presented with the complaint of anorexia, persistent recumbency and loss of body weight which lasted for few days. On clinical examination, the affected dog revealed rise in temperature (105°F), lachrimation, anaemia, unilateral corneal opacity, swelling of throat, enlargement of lymph nodes and severe loss of bodily condition. Wet smear, thin blood smear and hematological parameters were observed. Motile trypanosomes were found in wet smear, while trypanosomes with a characteristic flagellum, kinetoplast and undulating membrane were present outside the red blood cells in the thin smear. The dog was successfully recovered with a single dose of diminazene aceturate. This report may provide a way forward to establish effective and safe therapeutic protocols for the control of canine trypanosomosis.

HUBUNGAN INTERPRETASI WHO (WORLD HEALTH ORGANIZATION) ANTROPOMETRI Z-SCORE DAN INFEKSI KECACINGAN PADA ANAK USIA 36 – 60 BULAN DI KOTA LHOKSEUMAWE

Masalah pada anak yang masih menjadi perhatian khusus yaitu masalah status gizi. Penentuan status gizi yang sering digunakan adalah dengan interpretasi WHO antropometri z-score. Selain penentuan status gizi, sangat penting mengetahui infeksi yang dialami oleh anak, salah satunya yaitu infeksi kecacingan. Penelitian ini bertujuan untuk mengetahui karakteristik dan hubungan interpretasi WHO antropometri z-score berdasarkan Berat Badan (BB) menurut usia dan infeksi kecacingan pada anak usia 36 – 60 bulan di Posyandu Pusong Kota Lhokseumawe. Penelitian ini merupakan deskriptif analitik dengan rancangan cross sectional. Sampel penelitian adalah anak di Gampong Pusong yang memenuhi kriteria inklusi dan eksklusi. Interpretasi menggunakan software WHO Anthro serta infeksi kecacingan dilakukan dengan pemeriksaan feses dengan teknik direct thin smear feces menggunakan mikroskop. Data dianalisis secara secara univariat dan bivariat. Hasil penelitian didapatkan anak usia 36-47 bulan sebanyak 44,7% serta usia 48 – 60 bulan sebesar 55.3%; 57,9% adalah perempuan, dengan rata – rata berat badan adalah 14±2.07 kg dan rata – rata tinggi badan adalah 92±6,71 cm. Interpretasi WHO antropometri z-score berdasarkan BB menurut usia diperoleh sebesar 60,5% dengan status gizi baik, 39,5% dengan status gizi kurang, serta tidak ditemukan gizi lebih dan gizi buruk. Sebanyak 26,3% positif mengalami infeksi kecacingan dengan 80% mengalami infeksi oleh cacing Ascaris lumbricoides serta 20% mengalami infeksi oleh cacing Ascaris lumbricoides + Trichuris trichiura. Hasil analisis bivariat didapatkan tidak terdapat hubungan antara interpretasi WHO antropometri z score berdasarkan BB menurut usia dengan infeksi kecacingan pada anak usia 36 – 60 bulan di Posyandu Gampong Pusong Kota Lhokseumawe.

2589. Two Cases of Congenital Babesiosis

Background Babesiosis is caused by Babesia microti and often transmitted via Ixodes scapularis. To the best of our knowledge, only 9 cases of vertical transmission have been reported. The spectrum of clinical presentation and optimal therapy for this population remains unknown. Methods Case 1 is a 4 week old female admitted with fever and irritability for 2 days. She was pancytopenic, with Hgb of 9.2 g/dL, Plt of 57 K/mm3, and absolute neutrophil count (ANC) of 500/mm3. Thin smear revealed 2.5% parasitemia. Mother was diagnosed with acute Lyme disease in the seventh month of pregnancy. Maternal serologies were positive for B. microti (IgM 1:100 and IgG >1:320). The infant received 1 PRBC transfusion and was treated with 10 days of atovoquone and azithromycin. Case 2 is a 5 week old female twin A admitted with 2 days of pallor, fatigue and poor feeding. Initial Hgb was 3.4 Gm/dL. Peripheral blood smear revealed many intraerythrocytic parasites consistent with B. microti. Thin smear obtained 24 hours after initiation of therapy showed 2% parasitemia. During hospitalization, the patient developed neutropenia with ANC nadir of 800/mm3. The patient required 4 PRBC transfusions and was treated with azithromycin and atovoquone for 10 days. The mother had an acute, self-limited febrile illness at 23 weeks gestation. At infant’s presentation, maternal serologies revealed negative B. microti IgM and positive IgG (1:160). Placental tissue from both twins was positive for B. microti DNA by PCR. Twin B was asymptomatic, had negative B. microti blood PCR, a negative B. microti IgM, positive IgG of 1:30 felt to represent transplacental maternal antibody, and did not require treatment. Results Both infants were successfully treated without relapse. Conclusion Congenital babesiosis is rare and may cause profound hematologic disturbances. We report 2 cases exhibiting neutropenia in addition to anemia and thrombocytopenia, supporting recent assertions by Wormser et al. that this is a common finding. In addition, Case 2 presented with a severe hemolytic anemia significantly worse than previously reported. Finally, we demonstrated successful treatment in neonates without exchange transfusion, even with severe anemia. Disclosures All authors: No reported disclosures.

A Survey of Trypanosome Species Circulating In Pigs In Three Selected Local Government District of Enugu State, Nigeria

A cross sectional survey of trypanosomosis of pigs was carried out in three randomly selected Local Government Areas (Nsukka, Udenu and Igbo Eze South) out of the six local government areas in Enugu North Senatorial Zone. Blood samples were randomly collected from a total of 1800 pigs from abattoirs and farms and were examined for trypanosomes parasites using standard parasitological methods: wet film, buffy coat and Giemsa-stained thin smear preparations. A total of 93 (5.2%) pigs were positive overall. Among the positive samples, Trypanosoma brucei was the predominant species (86.0%), followed by T. congolense (13.9%). The different relative isolation rate for Nsukka, Igbo-Eze South and Udenu Local government areas were 14 (4.7%), 14 (4.7%), and 19 (4.0%) respectively in the dry season, and 17 (5.7%), 17 (5.7%) and 12 (6.3%) in the rainy season respectively. The trypanosome prevalence in males from Nsukka, Igbo-Eze South and Udenu local governments were 6.5%, 6.2% and 4.2% respectively, while report in female were 3.8%, 4.1% and 5.9% in the three local governments respectively. There was no significant variations (p<0.05) with seasons, sex and age groups. It was therefore concluded that porcine trypanosomosis is still endemic in Enugu North Senatorial Zone with T. brucei being the most incriminated as the cause of the disease in the zone.

Inactivating Zaire Ebolavirus in Whole-Blood Thin Smears Used for Malaria Diagnosis

Malaria is an important mimic or coinfection in potential Ebolavirus disease patients. Here, we evaluated the efficacy of the 100% methanol-inactivating Zaire Ebolavirus Makona variant for malaria thin-smear preparation. We determined that 100% methanol completely inactivated the virus after 15 min.