Prevalence and severity of gastro-intestinal parasites in buffalo calves at Sylhet division of Bangladesh

Abstract

Gastrointestinal (GI) parasites are one of the most widely recognized health problems of buffalo calves in Bangladesh. The study was undertaken to investigate the prevalence of GI parasitic infestation in buffalo calves at Sylhet division of Bangladesh. Fecal samples of 200 buffalo calves under 1 year of age (76 males and 124 females) were collected from Sylhet, Maulvibazar, Habiganj, and Sunamganj districts, and from each of the four regions, 50 samples were collected. Samples were examined through simple flotation, sedimentation, and McMaster techniques. The overall prevalence of GI parasites in Sylhet division was 65.5%. Among the samples, the highest prevalence of GI parasite was found in Sylhet (78%), followed by Sunamganj (68%), Maulvibazar (66%), and Habiganj (50%) districts. Among the identified GI parasites, the highest prevalence was observed in case of Neoascaris vitulorum (26.5%). The highest prevalence of GI parasite was observed in summer (69.84%), followed by rainy (69.62%) and winter (55.17%) seasons. Males and females were almost equally susceptible to GI parasitic infestation. Older calves (9–12 months, 77.14%) were most vulnerable to GI parasites. Egg per gram (EPG) of feces was additionally counted in the current study to know the severity of infestation. The range of EPG varied from 50 to 900 among the identified parasites. Among them, the highest count was found in the case of Neoascaris vitulorum (100–900). It may be concluded that aggregated policies and actions need to be taken to control the GI parasitic infestation in buffalo calves at Sylhet Division and elsewhere in developing countries like Bangladesh.

Introduction

Buffalo is one of the most important domesticated animals in Bangladesh which serves as a source of milk, meat, manure and drought power. It has a significant contribution in Gross Domestic Product (GDP) in terms of milk, meat, and skin, representing about 27.0%, 23.0%, and 28.0%, respectively to the entire output from the livestock sector in Bangladesh (Islam et al. 2018). In certain parts of Bangladesh, particularly in the hilly areas and low lying swampy lands, the farmers are more reliant on buffaloes than cattle for their livelihood due to lengthier working life of buffalo than that of cattle, typically over 17 years and up to 25 years of age (Fischer 1976). Buffaloes of Sylhet region are usually of swamp type rather than other regions of Bangladesh (Rahman et al. 2015). The rural poor farmers of this region usually rely on the buffaloes mostly for draught and meat purposes. Most of the day, the buffaloes were nibbling in the haor region, grazed in the pasture land, gathered in paddy fields, alongside the road, and satisfied their physiological requirements (Rahman et al. 2015). During that time, they might be infested with various parasites by ingestion as the low lying sloppy land and the stale water in haor areas mostly harbor the intermediate hosts and infective stages of parasites. The problem is mistreated because of its chronic and insidious nature (Sanyal 1998). The gastro-intestinal (GI) tracts of animals harbor an assortment of helminths, which cause clinical and subclinical parasitism. Helminths adversely affect animals’ health status, which may be a cause of economic losses to the buffalo industry. Helminths have been noted as a significant constraint to ruminant productivity on a clinical and subclinical level (Rahman et al. 2017). Parasitic diseases are emphasized for their pathogenicity and economic importance in animals by the experts both from the government and non-government organizations. The GI worms live in the digestive system in a larger organism or host (Rahman et al. 2017). Parasitic diseases are more significant in buffaloes than other infectious diseases (Roy et al. 2016).

GI parasites cause impaired digestion and affect the absorption of minerals, particularly calcium and phosphorus (Speedy 1992). GI parasitic infestations are common in buffaloes, particularly calves, causing considerable global economic losses to the buffalo industry and farming communities due to the mortality in infected young animals and reduced weight gain (Jyoti et al. 2016). Typical clinical manifestations perceived by the infected buffalo calves include anorexia, abdominal pain, dehydration, constipation or diarrhea, weight loss or poor weight gain, and a butyric smell on the breath (Li et al. 2015). Difference in agro-climatic conditions, animal husbandry practices, and pasture management largely determine the incidence and severity of different parasitic diseases in a particular region (Rahman et al. 2017). However, in developing countries like Bangladesh, minimal distributed data and information on the epidemiological aspects of helminth infestations in buffalo calves exist. Epidemiological patterns of parasitic diseases in different agro-climatic zones of the country would give a premise to develop key strategic control measures against the maladies. Research information is hardly accessible concerning the transmission of GI parasites of swamp type buffaloes in Bangladesh (Mamun et al. 2011; Biswas et al. 2014; Saha et al. 2014). Most of the rural people of Sylhet region in Bangladesh are living under the poverty line. They are directly involved in buffalo production which has been greatly hampered due to the detrimental effects of various GI parasites. Though epidemiological data are important for controlling GI parasites of buffalo calves, limited data are available on GI parasitic infestations in buffalo calves in Sylhet region of Bangladesh. Therefore, the present study was undertaken to determine the prevalence and severity of GI parasites in buffalo calves at Sylhet division of Bangladesh.

Materials and methods

Study area and study period

The study was conducted at Sylhet, Maulvibazar, Habiganj, and Sunamganj districts under Sylhet (marshy & hilly) division of Bangladesh from March 2018 to February 2019. The sampling was performed in three seasons according to the seasonal clarification of Bangladesh such as summer (March-June), rainy (July-October), and winter (November-February).

Study population and sample collection

A total of 200 fecal samples were collected randomly from individual buffalo calves from 10 different buffalo farms in each district and 5 samples were collected from each farms. Buffalo calves were divided into three age groups (1–4 months, 5–8 months, and 9–12 months) of either sex (males 76 and females 124) during the study period. All the samples were directly collected from the rectum or from the fresh fecal mass immediately after defecation of both diseased and apparently healthy buffalo calves. After collection, samples were moved to the laboratory of the Department of Medicine, Faculty of Veterinary, Animal and Biomedical Sciences, Sylhet Agricultural University (SAU), Sylhet, Bangladesh with maintaining proper cool chain.

Processing and examination of fecal samples

Specimens were kept in a refrigerator at 4 °C for some delayed examination. Nematodes eggs were identified by the floatation technique of fecal sample examination in saturated NaCl solution, and trematodes eggs were examined by the sedimentation technique, and finally, positive cases were counted through the McMaster technique. The degree of severity of infestation was determined through egg per gram (EPG) count, as described by Soulsby (1968).

The animals were then categorized as lightly, moderately, and severely (massively) infected according to the EPG of feces counts. EPG of feces counts ranged from 100–250, >250–500, and > 500 were considered as light, moderate and massive infestation, respectively.

Coproculture

Faecal samples were thoroughly crumbled before being mixed with sufficient vermiculite, using nonporous stampers, to a depth about 5 cm in wide mouthed glass jars with approximately 1 L capacity. A hole was left in the center of the culture by holding a stamper vertically in the center of the jar. The cultures were moistened sufficiently to ensure that it did not dry out during incubation period but not waterlogged. The jars were incubated in the dark at 26–28 °C for 14 days to ensure L3 development. All samples were then processed by Baermann technique by placing the larval culture directly in conical sedimentation glasses filled with tap water and left for 1–2 h. To stimulate larval migration, tap water was first heated until approximately 50 °C overnight (van Wyk and Mayhew 2013; Sengupta et al. 2016; Purwati et al. 2017).

Identification of the larvae:

Larval suspensions were dropped onto microscope slides. Larvae were stained with Lugol solution (5 g iodine crystals and 10 g potassium iodide in 100 mL distilled water). To preserve the shape of the larvae, 2% formalin was added. Tubes containing larvae were heated gently with Bunsen burner. By this step, larvae were dead but in preserved shape and stained well to ease the structure identification. Identification of strongyle larvae was based on the observation of the characteristic of the anterior part, sheet tail extension (STE) length (the thin shape extension of the sheath caudal), full body length and esophageal type under the light microscope (van Wyk and Mayhew 2013; Purwati et al. 2017).

Statistical analysis

The data were analyzed by using Microsoft Excel 2010 and Statistical Package for Social Science (SPSS) version 20. Results were expressed in percentage with P-value and significance was determined when P < 0.05.

Results and discussion

The findings of the present study showed that the overall prevalence of GI parasites in buffalo calves was 65.5% (Table 1). Among the GI parasites, the prevalence of Fasciola gigantica was revealed 14% individually in Sylhet Division while (Saha et al. 2014) detailed that fasciolosis in buffalo was 26.17 at Barisal in Bangladesh, which was almost like the detections at Sylhet Sadar but higher than the prevalence at Maulvibazar, Habiganj and Sunamganj region. The overall prevalence of Paramphistomum cervi was 9% in Sylhet division while (Saha et al. 2014) detailed that fasciolosis in buffalo was 60.75 at Barisal, Bangladesh, which was higher than the current findings. The findings of the present study showed a higher prevalence (26.5%) of Neoascaris vitulorum in buffalo calves which were found individually in Sylhet Division. Bhutto et al. (2002) recorded 33% prevalence of Neoascaris vitulorum in buffalo calves in Tandojam, Pakistan, which was higher than the present findings. Saha et al. (2014) found a 2% prevalence of Trichostrongylus sp., which was almost similar to the present finding, whereas 4% prevalence was recorded in all the Sylhet Sadar, Maulvibazar, Sunamganj, and no prevalence was recorded in Habiganj district. Jyoti et al. (2014) observed that the prevalence of Strongyloides sp. was 28.45%, which was much higher than the current study that is 6% overall prevalence in Sylhet division. In this investigation, the overall prevalence of Balantidium coli was 3% in Sylhet division, which was much lower than the investigation of (Roy et al. 2011) who reported 45.03% prevalence in buffalo at Mymensingh division in Bangladesh. (Biswas et al. 2014), who stated 7% prevalence of Eimeria spp in Bhola district, which is lower than the overall prevalence of 20% in Sylhet division in the present study.

Table 1.

Prevalence of gastro-intestinal parasitic infestation in buffalo calves at Sylhet division

Area	Sylhet (%)	Maulvibazar (%)	Habiganj (%)	Sunamganj (%)	Overall (%)	P-value
Name of the Parasites	(N = 50)	(N = 50)	(N = 50)	(N = 50)	(N = 200)
Fasciola gigantica	10 (20)	7 (14)	6 (12)	5 (10)	28 (14)	0.007(**)#
Paramphistomum cervi	7 (14)	5 (10)	2 (4)	4 (8)	18 (9)
Neoascaris vitulorum	17 (34)	12 (24)	10 (20)	14 (28)	53 (26.5)
Trichostrongylus sp.	2 (4)	2 (4)	0 (0)	2 (4)	6 (3)
Strongyloides sp.	3 (6)	2 (4)	4 (8)	3 (6)	12 (6)
Balantidium coli	2 (4)	0 (0)	2 (4)	2 (4)	6 (3)
Eimeria sp.	13 (26)	10 (20)	7 (14)	10 (20)	40 (20)
Subtotal	39+ (78)	33+ (66)	25+ (50)	34+ (68)	131+ (65.5)

P < 0.05 is significant, ⁺ = Total no. of animals affected is less than the summation of individual infestation because same animal was infected by more than one type of gastro-intestinal parasites, ^#variance of the prevalence of parasitic infestations among four districts

In this study, it was observed that the prevalence of GI parasites was somewhat higher in females (80, 55.1, and 69.6%) than in males (75%, 42.8%, and 64.7%) in Sylhet Sadar, Habiganj and Sunamganj district respectively (Table 2). These findings supported the previous report of Bhutto et al. (2002), who revealed a marginally higher prevalence (48.30%) of helminths in females than in males (45.12%) in buffalo calves in Pakistan. Yet, in Maulvibazar district, a somewhat higher prevalence was found in males (66.6%) than females (65.6%), which were supported by Raza et al. (2007) who stated the higher prevalence of GI parasites in male buffaloes than females in Pakistan.The present study also revealed that the prevalence of GI parasites in calves between 9 and 12 months age group were 93.3%, 81.2%, 66.7%, and 71.4% respectively in Sylhet, Maulvibazar, Habiganj and Sunamganj districts. Calves of this age group (9–12 months) were more susceptible to GI parasitic infestations than the age group of 5–8 months which were accounted as 93.3%, 73.3%, 47.6%, and 64.7% respectively in Sylhet, Maulvibazar, Habiganj and Sunamganj districts. Calves of 1–4 months of age were comparatively less susceptible to GI parasitic diseases (Table 3).

Table 2.

Sex wise prevalence of gastro-intestinal parasitic infestations in buffalo calves at Sylhet division

Area	Sex	F. gigantica	P. cervi	N. vitulorum	T. sp.	S. sp.	B. coli	E. sp.	Sub total (%)	P-value
Sylhet (%)	Male (N = 20)	4 (20)	3 (15)	8 (40)	2 (10)	1 (5)	1 (5)	5 (25)	15+ (75)	0.36 (NS)#
	Female (N = 30)	6 (20)	4 (13.3)	9 (30)	0 (0)	2 (6.6)	1 (3.3)	8 (26.6)	24+ (80)
Maulvibazar (%)	Male (N = 18)	3 (16.6)	3 (16.6)	5 (27.7)	1 (5.5)	1 (5.5)	0 (0)	4 (22.2)	12+ (66.6)	0.03(*)
	Female (N = 32)	4 (12.5)	2 (6.2)	7 (21.8)	1 (3.1)	1 (3.1)	0 (0)	6 (18.7)	21+ (65.6)
Habiganj (%)	Male (N = 21)	3 (14.2)	1 (4.7)	3 (14.2)	0 (0)	1 (4.7)	1 (4.7)	4 (19.0)	9+ (42.8)	0.55 (NS)
	Female (N = 29)	3 (10.3)	1 (3.4)	7 (24.1)	0 0	3 (10.3)	1 (3.4)	3 (10.3)	16+ (55.1)
Sunamganj (%)	Male (N = 17)	2 (11.7)	3 (17.6)	5 (29.4)	1 (5.8)	2 (11.7)	1 (5.8)	3 (17.6)	11+ (64.7)	0.19 (NS)
	Female (N = 33)	3 (9.0)	1 (3.0)	9 (27.2)	1 (3.0)	1 (3.0)	1 (3.0)	7 (21.2)	23+ (69.6)

N = Number of animals examined, P < 0.05 is significant, + = Total no. of animals affected is less than the summation of individual infestation because same animal was infected by more than one type of gastro-intestinal parasites, #variance among the parasitic prevalence between male and female, NS = Not significant, P > 0.05, #= Variance of the prevalence of parasitic infestations between male and female calves; F. gigantica = Fasciola gigantica, P. cervi = Paramphistomum cervi, N. vitulorum = Neoascaris vitulorum, T. sp.= Trichostrongylus sp., S. sp.= Strongyloides sp., B. coli = Balantidium coli, E. sp.= Eimeria sp

Table 3.

Age-wise prevalence of gastro-intestinal parasitic infestations in buffalo calves at Sylhet division

Area	Age	F. gigantica	P. cervi	N. vitulorum	T. sp.	S. sp.	B. coli	E. sp.	Sub total (%)	P-value
Sylhet (%)	1 day-4 months (N = 20)	0 (0)	0 (0)	9 (45)	0 (0)	0 (0)	0 (0)	4 (20)	11+ (55)	0.13 (NS)#
	5–8 months (N = 15)	2 (13.3)	1 (6.67)	5 (33.3)	1 (6.6)	1 (6.6)	0 (0)	7 (46.6)	14+ (93.3)
	9–12 months (N = 15)	8 (53.3)	6 (40)	3 (20)	1 (6.6)	2 (13.3)	2 (13.3)	2 (13.3)	14+ (93.3)
Maulvibazar (%)	1 day-4 months (N = 19)	0 (0)	0 (0)	7 (36.8)	0 (0)	0 (0)	0 (0)	3 (15.7)	9+ (47.3)	0.16 (NS)#
	5–8 months (N = 15)	2 (13.3)	2 (13.3)	3 (20)	1 (6.6)	0 (0)	0 (0)	4 (26.6)	11+ (73.3)
	9–12 months (N = 16)	5 (31.2)	3 (18.7)	2 (12.5)	1 (6.2)	2 (12.5)	0 (0)	2 (12.5)	13+ (81.2)
Habiganj (%)	1 day-4 months (N = 15)	0 (0)	0 (0)	7 (46.6)	0 (0)	0 (0)	0 (0)	2 (13.3)	5+ (33.3)	0.48 (NS)#
	5–08 months (N = 17)	0 (0)	0 (0)	2 (11.7)	0 (0)	2 (11.7)	1 (5.8)	4 (23.5)	8+ (47.0)
	9–12 months (N = 18)	6 (33.3)	2 (11.1)	1 (5.55)	0 (0)	2 (11.1)	1 (5.5)	1 (5.55)	12+ (66.6)
Sunamganj (%)	1 day-4 months (N = 12)	0 (0)	0 (0)	9 (75)	0 (0)	1 (8.3)	0 (0)	1 (8.33)	8+ (66.6)	0.96 (NS)#
	5–8 months (N = 17)	1 (5.8)	1 (5.8)	3 (17.6)	1 (5.8)	1 (5.8)	0 (0)	6 (35.2)	11+ (64.7)
	9–12 months (N = 21)	4 (19.0)	3 (14.2)	2 (9.5)	1 (4.7)	1 (4.7)	2 (9.5)	3 (14.2)	15+ (71.4)

N = Number of animals examined, ⁺ = Total no. of animals affected is less than the summation of individual infestation because same animal was infected by more than one type of gastro-intestinal parasites, # = Variance of the prevalence of parasitic infestations among different age group of calves^, NS = Not significant, P > 0.05

The findings of this study were supported by (Biswas et al. 2014), who reported adult animals (88.81%) were more susceptible to GI parasitic infestation than young animals (83.45%) and calves (68.05%). The findings of the present study were not in agreement with Mamun et al. (2011), and Raza et al. (2007), who reported that higher prevalence of helminths infestation was found in young animals compared to adults in buffalo. Seasonal fluctuation of the year had no significant ( P > 0.05) effect on the prevalence of gastro-intestinal parasitic infection in buffalo calves (Table 4). A relatively higher prevalence with GI parasites was observed in the rainy season (84.2% and 71.4%), followed by summer (75% and 11%) and winter (73.3% and 53.8%) seasons in Sylhet Sadar and Maulvibazar district, respectively. Our findings were supported by Mamun et al. (2011) who stated that higher infestation of GI parasites were found in the rainy season (71.70%), followed by summer (58.90%) and winter (52.27%) seasons. However, in Habiganj district higher prevalence was found in summer (69.2%) followed by rainy (55%) and winter (29.4%) which was sustained by (Biswas et al. 2014) who expressed that the prevalence of GI parasites was minimal higher in summer (84.62%) followed by rainy (83.62%) and winter (81.16%) seasons, and the prevalence in Sunamganj district was the highest in winter (69.2%) season followed by rainy (68.4%) and summer (66.6%) seasons which was in contrast to every single other finding. The degree of severity of the GI parasitic infestation was assessed by the Egg per Gram (EPG) of feces of buffalo calves (Table 5). The range of EPG varied from 50 to 900 among the identified parasites. Among them, the highest count was found in the case of Neoascaris vitulorum (100–900).

Table 4.

Seasonal prevalence of gastro-intestinal parasitic infestation in buffalo calves at Sylhet division

Area	Season	F. gigantica	P. cervi	N. vitulorum	T. sp.	S. sp.	B. coli	E. sp.	Sub total (%)	P-value
Sylhet (%)	Summer (N = 16)	3 (18.7)	2 (12.5)	10 (62.5)	0 (0)	0 (0)	1 (6.2)	2 (12.5)	12+ (75)	0.83 (NS)#
	Rainy (N = 19)	5 (26.3)	4 (21.0)	5 (26.3)	2 (10.5)	2 (10.5)	1 (5.2)	3 (15.7)	16+ (84.2)
	Winter (N = 15)	2 (13.3)	1 (6.6)	2 (13.3)	0 (0)	1 (6.6)	0 (0)	8 (53.3)	11+ (73.3)
Maulvibazar (%)	Summer (N = 16)	2 (12.5)	1 (6.2)	8 (50)	1 (6.2)	1 (6.2)	0 (0)	2 (12.5)	11+ (68.7)	0.56 (NS)
	Rainy (N = 21)	4 (19.0)	3 (14.2)	3 (14.2)	1 (4.76)	1 (4.76)	0 (0)	3 (14.2)	15+ (71.4)
	Winter (N = 13)	1 (7.6)	1 (7.6)	1 (7.6)	0 (0)	0 (0)	0 (0)	5 (38.4)	7+ (53.8)
Habiganj (%)	Summer (N = 13)	2 (15.3)	0 (0)	6 (46.1)	0 (0)	1 (7.6)	0 (0)	1 (7.6)	9+ (69.2)	0.16 (NS)
	Rainy (N = 20)	4 (20)	2 (10)	3 (15)	0 (0)	2 (10)	1 (5)	3 (15)	11+ (55)
	Winter (N = 17)	0 (0)	0 (0)	1 (5.88)	0 (0)	1 (5.88)	1 (5.88)	3 (17.6)	5+ (29.4)
Sunamganj (%)	Summer (N = 18)	2 (1.11)	1 (5.5)	10 (55.5)	0 (0)	1 (55.5)	1 (55.5)	1 (55.5)	12+ (66.6)	0.89 (NS)
	Rainy (N = 19)	2 (10.5)	2 (10.5)	4 (21.0)	1 (5.2)	1 (5.2)	0 (0)	2 (10.5)	13+ (68.4)
	Winter (N = 13)	1 (7.6)	1 (7.6)	0 (0)	1 (7.6)	0 (0)	1 (7.6)	7 (53.8)	9+ (69.2)

N = Number of animals examined, ⁺= Total no. of animals affected is less than the summation of individual infestation because same animal was infected by more than one type of gastro-intestinal parasites, # = Variance of the prevalence of parasitic infestations among different seasons, NS = Not significant, P > 0.05

Table 5.

Summary statistics (range, mean ± SE) of EPG (Egg per gram of feces) of gastro-intestinal parasitic infestations in buffalo calves at Sylhet division

Area	EPG	F. gigantica	P. cervi	N. vitulorum	T. sp.	S. sp.	B. coli	E. sp.
Sylhet (%)	Range	50–250	50–200	100–900	50–100	50–150	50–150	50–400
	Mean ± SE	120.0 ± 21.34	100.0 ± 21.82	**500 ± 61.24	75.00 ± 25.0	100.0 ± 28.87	100.0 ± 50.00	234.6 ± 26.18
Maulvibazar (%)	Range	50–250	50–200	100–700	50–150	50–100	100-00	100–300
	Mean ± SE	128.6 ± 28 0.5	120.0 ± 25.50	*287.5 ± 49.28	100.0 ± 50.0	75.00 ± 25.00	100.0 ± 00	222.2 ± 25.15
Habiganj (%)	Range	50–200	100–150	100–450	100-00	50–150	50–100	50–250
	Mean ± SE	108.3 ± 23.86	125.0 ± 25.00	227.8 ± 40.06	100.0 ± 0.00	87.50 ± 23.94	75.00 ± 25.00	142.9 ± 25.42
Sunamganj (%)	Range	50–250	100–200	100–550	100–150	50–100	50–100	50–200
	Mean ± SE	150.0 ± 35.36	137.5 ± 23.94	*289.3 ± 38.93	125.0 ± 25.0	66.67 ± 16.67	75.00 ± 25.00	135.0 ± 18.33

Degree of severity

100–250 = Light, 250–500 = Moderate, > 500 = Massive Infestation, (Soulsby, 1982; Urquhart et al., 1996; Maff, 1997), * = moderately infected, ** = severely infected

Conclusions

The results of the present investigation indicated that the prevalence of gastro-intestinal parasites in buffalo calves was very common irrespective of age and sex of the buffalo calves and seasons of the year. Based on the severity of infestation, Neoascaris vitulorum was identified as the GI parasite that affected buffalo calves more severely. It is suggested that proper strategies and preventive measures should be taken to control the infestation of GI parasites in buffalo calves in Sylhet division and elsewhere in Bangladesh.

Author contribution

MMR, MMR and AH provided the research proposal, IA, JA, PMD, SDN, TA, MRA, BD, SA and MQC collected the samples and performed laboratory works. MMR and MMR supervised the project. MMR was a scientific and lab diagnostic advisor to the project. IA analyzed the data. The manuscript was written by AA, finally revised by MMR. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

Ethical approval for the present study was duly obtained from and approved by the Institutional Animal ethics and Research committee of Sylhet Agricultural University, Sylhet, Bangladesh.