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I. PHOTOZO~L., 2, 19-20 (1955) Entamoeba bubalus, n.sp., from Carabao 19 GLENN A. NOBLE Califurnia Sfnte Polytechnic College, Sun Luis Obispo, California ~- c__ . SUMMARY. 'lwel\e carabao in the Philippines were found to be lightly infected with intestinal amoebae. Trophic forms (12 microns in diameter) of this protozoan possessed a definite ectoplasm and a homogeneous endoplasm. They were found only in stained preparations. The nucleus was similar to that of the cyst. All unconcentratcd fecal smears contained at least a few cysts (8 microns in diameter). In these forms the cytoplasm usually contained a large vacuole and one or more irregular chromatoidal bodies. The nucleus (2.6 microns in diameter) possessed a pronounced, deeply staining, uniform peripheral ring and a large irregular endosome. There was no periendosomal ring, This amoeba is designated as E77lnnto~bn bzlbnllc.r, 11.s~. URIhG THE academic year 1953-1954 the author had an opportunity, as a Lecturer at the Univer- sity of the Philippines under the Fulbright program, to continue a study of intestinal amoebae of domestic mammals Almost all of the large animals examined, as in other countries(3). contained amoebic cysts. This report is limited to the results of a study of fifteen water buffalo, or carabao (Bububs bubulis) from sev- eral islands of the Philippine group. Twelve of these animals were infected with amoebae. the cytoplasm. The large endosome, 2 microns in diameter, was characteristic. This structure seemed to lie a group of several closely adjacent granules. Twenty-three cysts (Figs. 2 and 3) were found. 'Their average diameter was 3 microns (range 5-9). All of th,em contained one or more vacuoles. Usually there was a single large vacuole which crowded all cell contents to the periphery. The cytoplasm was made up of fine granules. Chromatoidal bodies were present MATERIALS AND P1ETHOL)S Fecal material was esamined a5 fresh as possible with saline and with iodine dilurnts. Samples were taken at random from the inass. Heidenhain's iron hematoxylin. using 2% phosphotungstic acid for destaining. was employed in making permanent slides. Outlines of the protozoa and their organelles were drawn with the aid of a camera lucida and were used as the basis for illus- trations. Measurements were taken with an ocular micrometer which was calibrated against a standard stagt micrometer for each magnification used. RESliLTS .-\lthuu~h most of the carabao harbored amoebae. infection was light in every infected animal. Kumbers of cysts per entire slide ranged from 0 to 6. I-sually there was only one or two. The blood of one of the host animals was infected with TQJ~UITOSO?PZO ~vansi and several others harbored a species of intestinal trichomonad. All carabao were afflicted with intes- tinal worms. They Only two motile forms (Fig. 1) were seen.

Entamoeba bubalus, n.sp., from Carabao

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Page 1: Entamoeba bubalus, n.sp., from Carabao

I . PHOTOZO~L., 2, 19-20 (1955)

Entamoeba bubalus, n.sp., from Carabao

19

GLENN A. NOBLE Califurnia Sfnte Polytechnic College, Sun Luis Obispo, California

~- c__ .

SUMMARY. 'lwel\e carabao in the Philippines were found to be lightly infected with intestinal amoebae. Trophic forms ( 1 2 microns in diameter) of this protozoan possessed a definite ectoplasm and a homogeneous endoplasm. They were found only in stained preparations. The nucleus was similar to that of the cyst. All unconcentratcd fecal smears contained at least a few cysts (8 microns in diameter). In these forms the cytoplasm usually contained a large vacuole and one or more irregular chromatoidal bodies. The nucleus (2.6 microns in diameter) possessed a pronounced, deeply staining, uniform peripheral ring and a large irregular endosome. There was no periendosomal ring, This amoeba is designated as E77lnnto~bn bzlbnllc.r, 11.s~ .

U R I h G THE academic year 1953-1954 the author had an opportunity, as a Lecturer at the Univer-

sity of the Philippines under the Fulbright program, to continue a study of intestinal amoebae of domestic mammals Almost all of the large animals examined, as in other countries(3). contained amoebic cysts. This report is limited to the results of a study of fifteen water buffalo, or carabao ( B u b u b s bubulis) from sev- eral islands of the Philippine group. Twelve of these animals were infected with amoebae.

the cytoplasm. The large endosome, 2 microns in diameter, was characteristic. This structure seemed to lie a group of several closely adjacent granules.

Twenty-three cysts (Figs. 2 and 3 ) were found. 'Their average diameter was 3 microns (range 5-9). All of th,em contained one or more vacuoles. Usually there was a single large vacuole which crowded all cell contents to the periphery. The cytoplasm was made up of fine granules. Chromatoidal bodies were present

MATERIALS A N D P1ETHOL)S

Fecal material was esamined a5 fresh as possible with saline and with iodine dilurnts. Samples were taken at random from the inass. Heidenhain's iron hematoxylin. using 2% phosphotungstic acid for destaining. was employed in making permanent slides. Outlines of the protozoa and their organelles were drawn with the aid of a camera lucida and were used as the basis for illus- trations. Measurements were taken with an ocular micrometer which was calibrated against a standard stagt micrometer for each magnification used.

RESliLTS

.-\lthuu~h most of the carabao harbored amoebae. infection was light in every infected animal. Kumbers of cysts per entire slide ranged from 0 to 6. I-sually there was only one or two. The blood of one of the host animals was infected with TQJ~UITOSO?PZO ~ v a n s i and several others harbored a species of intestinal trichomonad. All carabao were afflicted with intes- tinal worms.

They Only two motile forms (Fig. 1) were seen.

Page 2: Entamoeba bubalus, n.sp., from Carabao

20 Entamoeba bubalus

in all cysts. Th,ese bodies were occasionally large with rounded ends s i rnihr to those of Entnriic~cbu histol?tic-o (Fig. 2 ) , but more often were small and irregular in size and shape (Fig. 3 ) . The nucleus averaqetl 2.6 microns (range 1.2 - 3.3) in diameter and was. in gen- eral, similar in appearance to that of the motile form. A distinct peripheral ring was usually seen. Chromatin distribution in the nuclear wall. however. varied. In some organisms there was practically none. in others there appeared a ring of dots. in still others there were a few isolated larger clumps. The endosome averaged 1.4 microns (range 0.8 - 1.6) in diameter. Often this structure appeared to b,e a cluster of four cranule.. There was no periendosomal ring.

DISCUSSION W D GENER.\L C’OYCLUSIONS

Earlier work on the genus Enturriocbn in domestic mammals threw doubt on the validity of assigning dif- ferent species names to thesc intestinal amoebae. ,L\ recent study of the amoebae of Philippine goats( 2 ) . however. indicated a new species from this host. In the

carabao the amoebae are likewise distinctive enough to be called a different species. The nucleus. especially. has constant diagnostic characteristics. Its heavy. usually uniform, outer ring and large prominent endo- some serve to distinguish this species. Compared to any random selection of twenty-three amoebic cysts from other domestic mammals. those from carabao showed remarkably little variation in structure. The distinctiveness of this amoeba and its apparent limita- tion to the carabao warrants its designation as Elztn- rnochn hirhnlus, n.sp. Cross-infection experiments. however. should be carried on. As Chen ( 1 ) and others have emphasized. host specificity is one of the best means of identifying species of all types of paraTites.

REFERENCES

1. Chtm. T. H . (1940). Morphological and developmental .tudirs (11 Paragonimus iloktsuensis with some remarks on other specie5 o i the genus (Trematodn : Troglotrematidae) . Lingnnn Sri. .I., 19. 4?9-5.10.

2 . Koble, G. .4., (19.54). Eiztaiiroeba dilinzani, n s p from Philippine goats. Philippine J . Sci., (in press).

3 . Soble, G . -4. & Noble, E. R. ( 1 9 5 2 ) . Entamoebae from i:lrm mamm;rl.-. .I. Pnrn.pitol.. 38. 571-.5q5.