Jarernsak Salakij

Hematologic, cytochemical, ultrastructural, and molecular findings of Hepatozoon‐infected flat‐headed cats (Prionailurus planiceps)

BACKGROUND The flat-headed cat (Prionailurus planiceps) is a small wild cat of Southeast Asia and is considered extremely endangered. Little is known about the hematologic values, blood cell morphology, or hemoparasites of this species in relation to other Felidae. OBJECTIVES The objective of this study was to report basic hematologic values and describe the light microscopic, cytochemical, and ultrastructural characteristics of blood cells in 2 wild-caught flat-headed cats. In addition, molecular analysis was done of a Hepatozoon organism found in the neutrophils of both cats. METHODS Blood samples were collected into EDTA from the cephalic vein. A CBC, manual differential count, manual reticulocyte count, cytochemical stains (Sudan black B [SBB], alpha-naphthyl acetate esterase [ANAE], and beta-glucuronidase), and scanning and transmission electron microscopy were done using standard methods. RESULTS HCT was slightly lower and reticulocyte counts and red cell distribution width were higher than the expected values for other species of cats. Hepatozoon organisms were found in the cytoplasm of neutrophils in both cats, but the number of infected neutrophils was very low (1%-2%). Neutrophils stained strongly positive for SBB, but were negative for ANAE and beta-glucuronidase. Hepatozoon-infected neutrophils were negative for SBB, but focally positive for ANAE and beta-glucuronidase. By transmission electron microscopy, gamonts of Hepatozoon sp were observed in neutrophils, and rarely free in plasma. Infected neutrophils had fewer specific granules and more mitochondria compared with noninfected neutrophils. PCR products of partial 18S rRNA revealed that the isolate of Hepatozoon in the flat-headed cats was closely related to that of the frog Hepatozoon sp. CONCLUSIONS These results add to our understanding of hematologic values and blood cell morphology in Hepatozoon-infected flat-headed cats as well as the molecular analysis of the Hepatozoon organism, and may be useful for the health management and evaluation of hemoparasitic disease in this species.

Molecular characterization of Anaplasma platys in a domestic cat from Thailand

A 5-year-old female cat was presented at Kamphaengsaen Animal Hospital for a complication of jaundice. Haematology revealed mild non-regenerative anaemia, lymphopenia, thrombocytopenia, hyper-proteinaemia and icteric plasma. The inclusion bodies in the platelets were detected in a stained blood smear. Partial sequences of the 16S rRNA gene from the isolate indicated that it had the closest relation to Anaplasma platys sequences from many dogs in Thailand and those obtained from GenBank. This is the first report of Anaplasma platys in a naturally infected domestic cat from Thailand.

Characterization of blood cells in the Leopard Cat (Prionailurus bengalensis)

BACKGROUND The Leopard Cat (Prionailurus bengalensis) is the most frequently encountered wild cat in most of Southeast Asia. Limited hematologic investigation exists for this species. OBJECTIVES The objectives of this study were to assess routine hematologic measurements and parameters and characterize the morphology, cytochemical staining, and ultrastructural features of blood cells in Leopard Cats. METHODS Blood samples were collected from 12 adult healthy captive Leopard Cats (7 males and 5 females). Complete blood counts were performed using an automated hematology analyzer and manual differential counts. Cytochemical staining (Sudan black B [SBB], peroxidase [PO], periodic acid-Schiff [PAS], alpha-naphthyl acetate esterase [ANAE], and beta-glucuronidase [BG]) and scanning and transmission electron microscopy were performed using standard methods. RESULTS Median (range) hematologic results were as follows: PCV 0.46 L/L (0.30-0.55 L/L), hemoglobin 136.5 g/L (100-183 g/L), WBC 9.0 x 10(9)/L (6.9-15.2 x 10(9)/L), band neutrophils 0.07 x 10(9)/L (0-0.30 x 10(9)/L), segmented neutrophils 2.9 x 10(9)/L (1.2-6.34 x 10(9)/L), lymphocytes 5.3 x 10(9)/L (2.7-8.1 x 10(9)/L), eosinophils 0.14 x 10(9)/L (0-0.73 x 10(9)/L), basophils 0/L (0-0.22 x 10(9)/L), and monocytes 0.08 x 10(9)/L (0-0.30 x 10(9)/L). Neutrophils stained strongly positive for SBB, PO, and PAS; lymphocytes had fine granular positivity for ANAE and BG; monocytes were weakly positive for ANAE and BG; and basophils were strongly positive for BG. Ultrastructurally, eosinophils contained many large rod-shaped granules with prominent crystalloid core structures, ribosomes, and mitochondria. Basophils contained many round to oval specific granules with homogeneous contents. Low number of basophils contained a few small vacuoles that usually were not detected by light microscopy. CONCLUSION These findings will facilitate interpretation of hematologic results for future investigative and diagnostic studies of this species.

Characterisation of blood cells in jungle cat, Felis chaus (Carnivora, Felidae)

The jungle cat (Felis chaus subsp. fulvidina) of Thailand is a protected animal. Haematological and morphological characteristics of blood cells in eight captive jungle cats (three males and five females) were investigated using complete blood cell counts, cytochemical stains and scanning and transmission electron microscopy. Haematological results were reported as mean, median and range (minimum–maximum). Neutrophils stained strongly positive for Sudan black B (SBB) and peroxidase (PO). Basophils were strongly positive for α-naphthyl acetate esterase (ANAE) and β-glucuronidase (BG) while eosinophils were negative for all cytochemical stains. Monocytes were negative for SBB and had fine granular reactivity for PO, ANAE and BG. The ultra-structure of eosinophils and basophils were different from that reported for other wild cats in Southeast Asia. The haematology, cytochemistry and ultra-structure of blood cells in the jungle cat may be useful in evaluating and monitoring the health of this species in captivity.

Haemoproteus in barn and collared scops owls from Thailand

The barn owl (BO) and the collared scops owl (CSO) are common nocturnal raptors throughout Thailand. Blood samples from 23 adult BOs and 14 CSOs were collected and processed for complete blood cell counts and parasite morphological examinations. Two Haemoproteus-positive samples were processed for ultrastructural observation. Polymerase chain reaction (PCR) analysis for a partial cytochrome b gene (cytb) from Haemoproteus was performed in all samples. Haemoproteus presence detected by light microscopy was lower than that detected by PCR (30.4% and 34.8%, respectively, in BO; and 50.0% and 78.6%, respectively, in CSO). Comparative hematology revealed that Haemoproteus-positive BOs had higher mean cell hemoglobin concentration, total leukocyte, absolute heterophil, basophil, and monocyte counts than Haemoproteus-negative BOs, but no significant differences between Haemoproteus-negative and -positive CSOs. Monocyte ultrastructure analysis revealed a role in the elimination of gametocytes. Morphologically, the Haemoproteus in 3 BOs and 6 CSOs were identified as H. noctuae, while that in 1 CSO was identified as H. syrnii. Phylogenetic analysis indicated the Haemoproteus spp. in 8 BOs and 7 CSOs were not closely related to H. noctuae or H. syrnii, and the cytb of 2 CSOs was that of H. syrnii. These results should be useful for study of Haemoproteus.