Veterinaria, Vol. 64, No. 2, 2015
Research - Full Paper
Diarrhoea in cattle caused by Buxtonella sulcata in
Sarajevo area
Jasmin Omeragić 1 *, Ćazim Crnkić 2
Abstract
Recent studies showed that Buxtonella sulcata (B. sulcata) may be a potential causative agent of periodical
recurrent diarrhoea of unknown etiology in cattle. This study aimed to investigate the prevalence and intensity
of infection of B. sulcata and its role in diarrhoea in cattle in Sarajevo area. A total of 412 faecal samples
were collected from cattle of different ages (189 young and 223 adults). The overall rate of infection was
27.2%, with a significant difference between young and adults (33.3% vs 21.9%). In animals infected with B.
sulcata diarrhoea was present in 57.1% young and 51.0% adults, with no significant difference between them.
The frequency of diarrhoea incidence increased proportionally to the intensity of infection with B. sulcata,
either young or adults, in a typical logarithmic trend in both age groups (R2 =0.97). In samples with more than
2000 cysts per 1g of faeces diarrhoea was found in almost 80% of adults, and in 100% of young animals.
The results obtained here indicate that young animals are more prone to the infection with B. sulcata, but
regardless of age the cattle similarly responds to the infection.
Keywords
cattle — protozoa — Buxtonella sulcata — diarrhoea — Sarajevo
1 Department
of Parasitology and Invasive Diseases of Animals, Veterinary Faculty, University of Sarajevo, Zmaja od Bosne 90, B&H
of Animal Nutrition, Veterinary Faculty, University of Sarajevo, Zmaja od Bosne 90, B&H
*Corresponding author: [email protected]
2 Department
Introduction
The first written document on B. sulcata was given by
Jameson (9). He reported heavy intestinal infection with
B. sulcata in 5 out of 8 cattle from Cambridge. The most
prominent character of this new ciliate was a dorsal ridge
running in a wide sweeping curve from one end of the
body to the other with a groove running down the middle.
Roundish oval cysts of this ciliate were 80 to 100 µm in
length by 60 to 80 µm in width.
Rees (14) studied morphology and behaviour of B. sulcata from cattle and Balantidium coli from pig. A ciliate
occurring in the large intestine in about 25% of the cattle
was detected in slaughtered animals by examination of the
faecal material in Syracuse watch glasses under the binocular microscope. Henriksen (7) in Denmark, examined fecal
samples of 762 cows and found cysts of B. sulcata in 547
(71.8%) cases, and Fox and Jacobs (5) from UK found cysts
in 44.6% of fecal samples. Hong and Youn (8) reported
prevalence of B. sulcata in 33.6% cattle in 1984 and 34.5%
in 1994. In infected cattle diarrhoea symptoms have been
present in 184 (69.1%) animals from 1984 and in 56 (55.5%)
animals from 1994. Wacker et al. (21) reported presence
of B. sulcata in cattle in Germany and clinical symptoms
accompanied by diarrhoea in 73% of infected animals. The
lowest incidence of B. sulcata in cattle (7.5%) in European
countries was found in Greece (3).
Tomczuk et al. (18) examined fecal samples of 116
dairy cattle in Poland from 19 farms where they observed
recurrent diarrhoea of unknown etiology. B. sulcata cysts
were found in 102 (87.9%) animals, and diarrhoea was
present in 35 (34.31%) animals examined. The authors also
found that the diarrhoea intensity was proportional with the
In Bosnia and Herzegovina (B&H) diarrhoea caused by
protozoan parasites in cattle, especially in young animals,
was mainly attributed to the parasite phylum Apicomplexa,
such as species of the genus Eimeria and Cryptosporidium
(13). In the past, investigation on protozoan phylum Ciliophora did not attract special attention. It is known that
some ciliates are localized in the gastrointestinal tract of
ruminants, but their role has not been fully clarified. There
is a general opinion that the protozoans in the gastrointestinal tract are involved in the process of digestion of plant
feeds as a commensal organism. However, high incidence
of infection in ruminants with Buxtonella sulcata (B. sulcata) and changes that it causes, points out that in some
cases this ciliate may be responsible for the incidence and
intensification of diarrhoea symptoms (18). Urman and
Kellky (20) and Skotarczak (15) suggested that B. sulcata
can lead to pH changes of large intestinal content of cattle.
Also, the multiplication of the parasite causes a cytotoxic
effect in the large intestine which is manifested as lesions
of the intestinal mucosa followed by secondary bacterial
infections.
Protozoan parasite B. sulcata (Jameson, 1926) has been
classified in the Kingdom: Protozoa, Phylum: Ciliophora,
Class: Kinetofragminophorea, Order: Trichostomatorida,
Family: Pycnotrichidae, Genus: Buxtonella (17). Predilection site of B. sulcata is in the large intestine (colon) of
cattle (11). Parasites of the phylum Ciliophora, including
B. sulcata developed cilia to move, which are actually short
growths of plasma.
50
Diarrhoea in cattle caused by Buxtonella sulcata in Sarajevo area — 51/54
B. sulcata infection intensity. With the infection intensity
of over 1000 cysts per gram (CPG) of feces, diarrhoea was
found in 90% of animals tested, and in all animals with more
than 2000 CPG. By improving zoohygienic conditions and
parenteral treatment with 5% and 20% glucose, vitamin
C and vitamin B, along with a balanced mineral-vitamin
dietary supplement, the number of cysts was drastically
reduced and the diarrhoea ceased in all treated animals.
In Iraq, Al-Saffar et al. (1) conducted examination of
120 cattle fecal samples in the city of Mosul. Infection with
B. sulcata was present in 29 animals (24.2%), and the cysts
were found in 27 of 86 (31.39%) animals with diarrhoea
symptoms. In dairy cows of Costa Rica, Jimenez et al. (10)
have found B. sulcata in 38%, and 21,6% of beef cattle.
Tung et al. (19) examined 310 fecal samples of cattle in
Taiwan where B. sulcata was found in 26 (8.4%) animals.
Ganai et al. (6) reported the incidence of B. sulcata in
20.9% bovines (n=374) of R.S. Pura, Jammu (India) where
cattle showed higher infection rate (23.6%) than the buffaloes (18.5%). The infection was higher in young animals
(33.1%) than the adults (13.9%). Animals with diarrhoeic
feces had higher infection rate (38.5%) than the animals
with normal feces (9,9%), suggesting the influence of B.
sulcata on the incidence of diarrhoea.
Only one isolated study on B.sulcata was conducted in
cattle in B&H (13) where 2019 fecal samples were examined and the cysts of B. sulcata were found in 464 samples
(22,9%). However, only 93 samples from Sarajevo area
were examined and 37 (39.8%) were positive, more than
double percentage when compared to north-western part of
the country (17.2%). Due to small number of samples from
the Sarajevo area it was difficult to conclude that incidence
of B. sulcata was so much different between investigated
regions. Therefore, the aim of this study was to investigate
presence on B. sulcata in a larger group of animals in Sarajevo area to obtain more correct data about the prevalence
of infection.
Material and Methods
Study area. Sarajevo, the capital of B&H and its largest urban, cultural, economic and transportation center, is located
almost in the geographical center of B&H (coordinates N
43◦ 52 0 000 , E 18◦ 250 000 , average altitude is 511 m). Based
on the data of the Federal Office of Statistics of B&H (16)
the climate for the area of Sarajevo is characterized by moderate and mild continental. The average annual temperature
is 9,5 ◦ C; precipitation is approximately 900 l/m2 .
Sample collection and examination. During the twoyear study (2012 and 2013) 412 fecal samples of cattle of
different age were collected. All the samples were taken
directly from the animal rectum, and were classified according to age and diarrhoea symptoms. Samples were
stored at 4 ◦ C into an isothermic box and sent to the Laboratory of Parasitology, Veterinary Faculty of Sarajevo, B&H.
Coprological examination was carried out using standard
sedimentation and flotation techniques. Determination of
cysts B. sulcata was conducted on the basis of characteristic
morphological features and measurement of the parasite’s
diameter (Figure 1).
Statistical analysis. The data were recorded in two
groups according to the age of the cattle: young (below 1
year of age) and adults (above 1 year of age). The fecal
samples were divided into two groups: non-infected and
infected. The feces of infected animals were further subdivided either as diarrhoeic and non-diarrhoeic, or as four
groups according to the number of cysts in 1 g (CPG) of
feces. Results were analyzed using Chi square test with a
p <0.05 considered as significant. Fisher’s exact test was
used in the case of 2∗2 tables when any expected frequency
was too low for Chi square test. Regression analysis was
performed to estimate the relationships between the frequency of diarrhoea incidence and intensity of infection
with B. sulcata. All statistics were performed using Minitab
15 statistical software (12).
Results
Coprological examination of cattle feces in this study showed the presence of B. sulcata cysts in 112 (27.2%) animals (Table 1). Significantly higher prevalence of infection
(p=0.010) was found in young compared to adults (33.3%
vs 21.9%). The infection was associated with diarrhoea
in more than a half of the infected animals, either young
(57.1%) or adults (51%), with no significant difference between them (p=0.519).
Table 1. Infection rate with B. sulcata and diarrhoea
incidence in infected animals
Animals with diarrhoea
(% of infected animals)
Young
189
63 (33.3)*
36 (57.1)
Adults
223
49 (21.9)
25 (51.0)
Total
412
112 (27.2)
61 (54.5)
*Significantly higher than in adults (p=0.010)
Age of cattle
Examined
Infected (%)
The increasing frequency of diarrhoea incidence in infected animals was proportional to the B. sulcata infection
intensity (Table 2) having a logarithmic trend (R2 =0.98; y =
45.963Ln(x) + 26.93) and the frequency was significantly
different depending on the infection intensity (p<0.001).
The relationship was evident even when viewed separately
for young (p<0.001) and adults (p=0.018), with the same
type of logarithmic trend for both age groups (R2 =0,97) (Table 3). Logarithmic growth of diarrhoea frequency was
described by equation y = 52.141Ln(x) + 25.99 or y =
39.711Ln(x) + 27.474, for young and adults, respectively.
However, separate analysis of each infection intensity in
Table 3 showed no significant differences between young
and adult animals.
Table 2. The effect of B. sulcata infection intensity on
frequency of diarrhoea incidence
Infection
Number of
Animals with
intensity (CPG) infected animals diarrhoea (%)*
<500
45
11 (24.4)
500 - 1000
28
18 (64.3)
1000 - 2000
21
16 (76.2)
>2000
18
16 (88.9)
Total
112
61 (54.5)
*Signicantly different depending on infection intensity (p< 0.001)
Diarrhoea in cattle caused by Buxtonella sulcata in Sarajevo area — 52/54
Table 3. The effect of B. sulcata infection intensity and the animal age on frequency of diarrhoea incidence
Young
Adults
Infection
Number of
Animals with
Number of
Animals with
Fisher’s p
intensity (CPG) infected animals diarrhoea (%)* infected animals diarrhoea (%)*
<500
25
6 (24)
20
5 (25)
1
500-1000
16
11 (68,8)
12
7 (58.3)
0.698
1000-2000
13
10 (76,9)
8
6 (75)
1
>2000
9
9 (100)
9
7 (77.8)
0.471
Total
63
36 (57.1)
49
25 (51)
0.569
*Signicantly different depending on infection intensity (young p<0.001; adults p=0.018)
Figure 1. Cyst of Buxtonela sulcata (Olympus, digital
camera, 40X)
Discussion and conclusion
The overall percentage of B. sulcata infection in cattle was
lower in this study in comparison to the percentages of
infection in Denmark, Britain, and South Korea (5, 7, 8),
but was higher compared with the results of similar studies
conducted in India, Greece, Iraq, and Taiwan (1, 3, 6, 19).
However, the obtained result are much lower if compared
with the results obtained for Sarajevo area in our previous
study, but still higher than those results for north-western
part of the country (13).
More than a half of the infected animals in the present
study showed diarrhoea symptoms which is similar to the
results obtained in our earlier work (13). Compared to
results reported here, the diarrhoea incidence was higher in
infected cattle in South Korea and Germany (8, 21), while
in Poland and Iraq have been lower (1, 18).
The diarrhoea incidence increased proportionally with
B. sulcata infection intensity (Table 2) what is in agreement with observations presented by Tomczuk et al. (18),
although in their study the diarrhoea incidence was even
higher in the animals with highest infection intensity. Specifically, the authors found diarrhoea 100% animals that have
had more than 2000 CPG, in 88.2%, animals with 1000 to
2000 CPG, 75% animals with 500 to 1000 CPG, and 16.3%
animals with less than 500 CPG. The current study also
showed that the same relationship exists between the diarrhoea incidence and B. sulcata infection intensity either in
young or adult animals (Table 3). Considering together with
the results shown in Table 1, we suggest that cattle similarly
respond to B. sulcata infection regardless of age. However,
in routine practice we still can expect more frequent incidence of diarrhoea caused by B. sulcata in young animals,
bearing in mind that they have much higher prevalence of
infection compared to adults as shown in this (Table 1) and
other studies (6).
The results obtained here, as well as the findings from
other studies (5, 8, 18, 20) suggest that at low infection
intensity (<500 CPG) a relatively small number of animals
showed clinical signs, such as diarrhoea. However, in animals with the infection intensity of >500 CPG diarrhoea
incidence was markedly increased, reaching 100% in cases
with the highest infection intensity (>2000 CPG).
Besides the number of cysts in the gastrointestinal tract,
there are many other factors influencing the course of buxtonelosis, including virulence of the pathogens, host susceptibility, environmental conditions, etc. Important for
the parasite-host impact are the pH changes in the colon.
Acidity stimulates growth and multiplication of parasites
and increases the cytotoxic effects on host tissue. Detrimental effect on the intestinal mucosa is exacerbated and
the resulting inflammatory lesions are suitable places for
the development of secondary bacterial infections that further complicate the pathological changes (18). It is known
that decrease in colonic pH may be a consequence of feeding regimen. When high concentrate diets are fed, some
starch escapes ruminal degradation and passes to the hindgut
where it undergoes fermentation resulting in higher concentration of volatile fatty acids (2). The acids decrease colonic
pH creating environment that is favorable for growth of
some pathogens. Low colonic pH also makes mucosal tissues more vulnerable, regardless of the pathogens action
(4). Finally, all feeding situations when intestinal pH is
moving toward acidic conditions can complicate protozoal
infection. Even when intestinal pH is not compromised,
increased infection with the protozoan parasites may result
in the accelerated passage of alimentary contents in the gut
causing disorders such as diarrhoea or poor condition of
animals (18). If the primary infection with B. sulcata is
not complicated by disruption of bacterial flora, then the
dietary changes and pH regulation in large intestine with
probiotics and symptomatic therapy would normally satisfy
therapeutic measures.
The results of this study showed that the prevalence of B.
sulcata infection in Sarajevo area is lower than previously
showed (13). Results confirm that young animals are more
prone to the B. sulcata infection but also indicate similar
infection responsiveness of cattle regardless of age.
Diarrhoea in cattle caused by Buxtonella sulcata in Sarajevo area — 53/54
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Diarrhoea in cattle caused by Buxtonella sulcata in Sarajevo area — 54/54
Dijareja goveda uzrokovana sa Buxtonella sulcata na području Sarajeva
Sažetak
Uvod
Ranija istraživanja su pokazala da Buxtonella sulcata (B.
sulcata) može biti potencijalni uzročnik periodičnih povratnih dijareja nepoznate etiologije u goveda. U BiH je do sada
provedeno samo jedno takvo istraživanje, a prisustvo cisti B.
sulcata utvrdeno je u 22,9% uzoraka fecesa. Medutim, na
području Sarajeva 39,8% uzoraka bilo je pozitivno, što je
dvostruko više u odnosu na sjeverozapadni dio zemlje, gdje
je stopa infestacije bila 17,2%. Zbog relativno malog broja
uzoraka sa područja Sarajeva (n=93), bilo je teško pouzdano zaključiti da su razlike izmedu dva navedena područja
uzorkovanja toliko različite. Cilj ovog rada bio je ispitati
učestalost i intenzitet zaraze sa B. sulcata na većem broju
uzoraka i utvrditi njenu ulogu u nastanku dijareje u goveda
na području Sarajeva.
Materijal i metode
Ukupno 412 uzoraka fecesa prikupljeno je od goveda različite dobi (189 mladih i 223 odraslih). Koprološka ispitivanja su provedena standardnim parazitološkim metodama
sedimentacije i flotacije. Determinacija cisti B. sulcata
provedena je na bazi morfoloških karakteristika i mjerenjem dijametra parazita.
Rezultati i interpretacija
Ukupna stopa infekcije bila je 27,2%, sa značajnom razlikom izmedu mladih i odraslih (33,3% nasuprot 21,9%).
U životinja zaraženih B. sulcata dijareja je bila prisutna u
57,1% mladih i 51% odraslih, bez značajne razlike medu
njima. Učestalost pojave dijareje povećavala se proporcionalno intenzitetu zaraze sa B. sulcata, bilo da su mlade
ili odrasle životinje u pitanju, u tipičnom logaritamskom
trendu rasta u obje dobne skupine (R2 = 0,97). U uzorcima
sa više od 2000 cista po 1 g fecesa, dijareja je ustanovljena
u gotovo 80% odraslih i 100% mladih životinja.
Zaključci
Dobiveni rezultati su pokazali da je stopa infekcije sa B.
sulcata na području Sarajeva manja od one koja je utvrdena
u ranijem istraživanju. Rezultati takoder ukazuju na to da
su mlade životinje više podložne infekciji sa B. sulcata, ali
i da goveda slično reagiraju na infekciju bez obzira na dob.