Sero-Prevenance of Coxiella burnetii and Associated risk factors in Small Ruminants from Central Nepal

doi:10.21203/rs.3.rs-4826760/v1

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Sero-Prevenance of Coxiella burnetii and Associated risk factors in Small Ruminants from Central Nepal

https://doi.org/10.21203/rs.3.rs-4826760/v1

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Q fever, also known as endemic zoonotic illness, is primarily thought to infect humans through ruminants. Coxiella burnetii, an obligatory intracellular bacterial pathogen with a global distribution, is the cause of Coxiellosis. A cross-sectional study was conducted with 88 goat serum samples from NARC, NGRP. Two (2) serum samples out of eighty eight (88) were confirmed to be positive for Coxiella burnetii, and the study found a prevalence of 2.273%. The risk factors of Q. fever such as age, physiological status, tick infestation, body condition score (BCS), history of abortion, health status, and stock source showed no significant correlation with finding. Further research on Q fever infection using interventional methods for control and prevention is therefore crucial from one health aspect.

Zoonotic

Q. fever

one health

risk factors

Query fever (Q-fever) is the one of most infectious infections caused by the bacteria, Coxiella burnetii, mainly caused by coxiellosis, a major worldwide occupational zoonotic illness. It is one of 13 worldwide priority zoonoses, which are highly contagious diseases caused by intracellular Gram-negative bacteria[1]. It is a highly contagious zoonosis prevalent in almost all mammals, affecting predominantly humans, cattle, sheep, and goats.[2]. The Centers for Disease Control and Prevention (CDC) classifies it as GROUP-B bioterrorism because of its relative morbidity, low fatality rate, and moderate to disseminatable nature, listed in the OIE notifiable illness database 2020, as well as the list of infectious agents capable of infecting various animal species[3][4]. It is distributed throughout the globe except in New Zealand and Antarctica where it is not reported[5]. The infectious dosage of Coxiella burnetii can be as low as one bacterium, which makes it "one of the highly contagious diseases," according to the International Livestock Research Institute (ILRI), due to its pathogen's infectious nature and the need for a biosafety level (BSL)-3 diagnostic approach[6].

Humans can become infected by eating contaminated milk, occasionally via tick bites, or by coming into close contact with diseased livestock while working on farms, Veterinary professionals, farm workers, and owners are more susceptible since they frequently handle small ruminants and contaminated materials.[7][8]. Clinical manifestations can vary and include self-limiting feverish infections with symptoms like exhaustion, irritation, general discomfort, myalgia, and joint pain, or more serious complications like neurologic signs, pneumonia, heart involvement, hepatitis, and persistent fatigue. About 60% of cases are asymptomatic[9]

Within the phagolysosomes of host monocytes and macrophages, C burnetii lives and reproduces. There are currently two types known: the large cell variation, which is found inside infected host cells, and the small cell variety, which is extracellular and present in milk, urine, and excreta, with the majority occurring in reproductive and placental fluids. The small variety exhibits resistance to both mild heat and conventional farm disinfectant, allowing the bacteria to live longer in contaminated materials and farm byproducts for a month in frozen meat, 10 months at the ambient temperature of 15–20 C, and greater than 40 months in skimmed milk at room temperature[1][10].

Study Site

A cross-sectional study was done between September and November, 2022 at Nepal Agriculture Research Council, National Goat Research Program (NGRP), NARC Bandipur, Tanahun District, Gandaki Province, Nepal(Fig. 1.). It is located in the hilly region at 27° 56' 21.83" North latitude and 84° 24' 3.22" East longitude, 141 km west of Kathmandu with an elevation of 812 m, annual rainfall is 2447 mm and an annual temperature is 16–27°C.

Flock Selection and Sampling Methods:

All female goats(88) at the research station were screened for annual routine screening, a total of 88 blood samples were collected from jugular vein and stored in EDTA tube (MED-VAC, Sparsh Mediplus, India), and transported to the station laboratory at 4°C, serum was separated with a pipette, stored in 1.5ml Eppendorf tube and stored at -20°C. ELISA test was performed according to the manufacturer’s protocol (ID Screen® Q Fever Indirect Multispecies, Innovative Diagnostic, France) at National Animal Health Research Division, Khumaltar, Kathmandu, Nepal.

Statistical analysis and interpretation:

Thus, the obtained results were entered in Ms-Excel. The prevalence rate, and statical significance according to age, physiological status, tick infestation, body condition score (BCS), abortion history, health condition, and stock source with lab results, compared and analyzed with Chi-Square (χ2) using IBM Statistical Package for Social Science (SPSS) software version 25 with significance level at 5%. (p < 0.05)

Table 1

Overall prevalence results of Coxiellosis
Total case	Positive case	Negative Case	Seropositivity %
88	2	86	2.273%

Table 2

Association between *Coxiell*a *burnetii* and the age of Goats
Variable	Positive Case	Negative Case	Seropositivity %	p-value
< 3 Years	1	34	2.857%	0.640 (NS)
> 3 Years	1	52	1.887%
Total	2(88)	86(88)	2.273%

Table 3: Association between Coxiella burnetii and the physiological status of Goats

Variable	Positive Case	Negative Case	Seropositivity %	p-value
Adults	1	61	1.613 %	0.506 (NS)
Young	1	25	3.846 %
Total	2(88)	86(88)	2.273 %

Table 4

Association between *Coxiell*a *burnetii* and tick infestation of Goats
Variable	Positive Case	Negative Case	Seropositivity %	p-value
Yes	1	12	7.692%	0.275 (NS)
No	1	74	1.333%
Total	2(88)	86(88)	2.273%

Table 5

Association between *Coxiell*a *burnetii* and body count score (BCS) of Goats
Variable	Positive Case	Negative Case	Seropositivity %	p-value
2.5	0	7	0	0.846 (NS)
2.5–3.5	2	79	2.470%
Total	2(88)	86(88)	2.273%

Table 6

Association between *Coxiell*a *burnetii* and abortion history of Goats
Variable	Positive Case	Negative Case	Seropositivity %	p-value
Yes	1	8	11.111%	0.195 (NS)
No	1	78	1.265%
Total	2(88)	86(88)	2.273%

Table 7

Association between *Coxiell*a *burnetii* and the health status of Goats
Variable	Positive Case	Negative Case	Seropositivity %	p-value
Healthy	1	74	1.333%	0.275 (NS)
Sick	1	12	7.692%
Total	2(88)	86(88)	2.273%

Table 8

Association between *Coxiell*a *burnetii* and stock source of Goats
Variable	Positive Case	Negative Case	Seropositivity %	p-value
Purchased	1	24	4%	0.490 (NS)
Reared	1	62	1.58%
Total	2(88)	86(88)	2.273%

This study shows 2.27% prevalence (Table. 1) in a blood sample of examined animals against C. burnetii. This was a routine seroprevalence study in this research station. There was non-significant association between age, physiological status, tick infestation, body condition score (BCS), abortion history, health condition, and stock source. The prevalence of C. burnetii was higher (2.857%) (Table. 2) in animals less than three (< 3) years old when compared to greater age (1.887%) (Fig. 2). Animals at young ages were more affected (3.846%) (Table. 3) than adults (1.613%)(Fig. 3). The level of prevalence was higher (7.692%) (Table. 4) in such animals which had tick infestation compared to those which hadn’t (1.333%)(Fig. 4). Similarly, prevalence was found higher in animals whose BCS score was higher (i.e. 2.5–3.5) (2.470%) (Table. 5) than in low BCS (2.5) score animals(Fig. 5). Animals with a history of previous abortion showed a higher prevalence (11.111%) (Table. 6) when compared with no abortion history (1.265%)(Fig. 6). In addition to abortion history, Animals with frequently sick and ill conditions had a higher prevalence(7.792%)(Table. 7) when compared with healthy(1.333%) animals(Fig. 7). Study found that animals purchased from outside of research station have higher prevalence (4%) (Table. 8) than those which were born and reared (1.58%) within the research station(Fig. 8).

The prevalence rate of 2.27% is closely in line with the study found 6.52% in Kavre and lower than the prevalence rate in Makawanpur 28.57% reported by a previous study[11]. It coincides study conducted on 184 cattle sera with 1.63% seropositivity from Rupandehi, Nepal[7]. Researchers from Bangladesh also reported a similar prevalence level of 3.33% in goat, 3.57% in cattle, and 9.52% in sheep[12]. Serological investigation in Saudi Arabia shows 44.6% prevalence followed by 36.8% prevalence in sheep[13]. However, these finding was comparatively lower than 11.3% among 300 blood samples from small ruminants from Punjab, Pakistan[14]. In our analysis, we did not find any statistically significant correlation. We were not able to discover any statistically significant association in our investigation. This doesn’t mean that there is no biological link, it was outside of the reach of our study.

A lot of infections that affect goats are zoonoses, such as C. burnetii, which can spread[15]. The current study and other studies carried out in different parts of the world have shown variations in the prevalence rates of C. burnetii infection in animals. These variations have been attributed to several factors, including the quantity and type of samples collected, the season, the location, the type of assay, potential variations among laboratories, testing protocols, and the standards used to define positive results.

This research proved the prevalence of Q. fever in small ruminants in the Mid-hill region of Nepal. This suggests that Nepal may be an important endemic site for Q. fever, further serological and PCR techniques are needed for disease surveillance and monitoring. Governmental regulatory authorities and related stakeholders should give proper awareness and information, repeated screening and investigation is essential for disease control. Safe animal trading for disease-free animals should implicated to minimize financial loss and the promotion of one health approach for disease control is essential.

Furthermore, studies and diagnoses of Q. fever in other livestock play an important linkage in studying disease transmission, control, outbreaks, and preventing health problems in Nepal.

Study approval: This study was approved and conducted at Nepal Agriculture Research Council, National Goat Research Program. Consent to conduct study and sample dispatch registration number: 103/2079/80

Funding: This Cross sectional study was supported by Nepal Agriculture Research Council, National Goat Research Program, NARC/Project/2080/81, Surveillance of common goat diseases in the outreach and research sites of NGRP, Bandipur, Tanahun. Unique Project Identifier (UPI) :4187681004 and Office LMBIS Code: 312413801.

Acknowledgment:

We would like to acknowledge all staff of National Goat Research Program, Bandipur, Tanahun.

Complecting interest:

The authors declares that they have no competing interests.

Author Contribution

R.G prepared the main manuscript, data analysis, and figures 1.P.S did an laboratory examination and Funding acquisitionS.A also prepared manuscriptAll authors reviewed the manuscript.

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No competing interests reported.

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You are reading this latest preprint version

Sero-Prevenance of Coxiella burnetii and Associated risk factors in Small Ruminants from Central Nepal

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Abstract

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Introduction

Materials and Methods

Study Site

Flock Selection and Sampling Methods:

Statistical analysis and interpretation:

Results

Discussion

Summary and Conclusion

Declarations

Author Contribution

References

Additional Declarations

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