Salmonella sp Contamination on Chicken Eggs and Duck Eggs at the Antasari Traditional Market, Banjarmasin, Indonesia

: Chicken eggs and duck eggs can be a source of transmission of Salmonellosis. Salmonella bacteria can penetrate the eggshell, or the egg's contents can become directly contaminated with Salmonella in the reproductive organs before the shell covers the egg. This research aims to analyze Salmonella sp in chicken and duck eggs at the Antasari Traditional Market in Banjarmasin, Indonesia. Samples were taken by purposive sampling with the criteria for cracked/broken eggs and intact eggs: 3 chicken eggs and three duck eggs from 6 traders so that the total sample was 24. For the identification of bacteria, microscopic tests and biochemical tests were used on the identification media. The results showed Salmonella sp in duck eggs and no Salmonella in chicken eggs. In all samples of chicken and duck eggs, 12.50% of Salmonella sp, 37.50% of Klebsiella sp, 12.50% of Proteus sp, and 37.50% of Pseudomonas sp were found. Suggestions for consumers are to pay more attention to the hygiene and sanitation of traders before buying eggs, choose clean eggs, store eggs in the refrigerator, and cook eggs until they are cooked.


INTRODUCTION
Nontyphoidal Salmonella is estimated to cause approximately 153 million cases of gastroenteritis and 57,000 deaths globally each year 1 .The disease caused by nontyphoidal Salmonella enterica is considered the most significant disease burden among enteric diseases 2 .
Several foods have been linked to cases and outbreaks of Salmonellosis 3 .Salmonellosis outbreaks linked to consumption of eggs and egg products accounted for the majority of outbreak-related illnesses in the United States at 2,422 illnesses 4 .Domingues et al.'s research (2012) stated that consumption of undercooked or raw eggs and poultry was found to be a risk factor for sporadic cases of Salmonellosis 5 .Laying chickens are the main reservoir of strains that cause Salmonellosis in humans, causing 42.4% of all cases of human infection and 95.9% of them caused by Salmonella enteritidis 6 .Salmonella enteritidis has been the leading cause of Salmonellosis in humans 7 , and this Salmonellosis is associated with eggs 8 Research in Indonesia by Riski L et al. (2023) on chicken eggs sold in traditional markets in Medan Johor District, the results showed that the average total colony of Salmonella sp. in Kwala Bekala Market was 1.45 x 105CFU/g in Johor Market it was 5.8

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x 104CFU/g.In Tikung Market it was 3.6 x 104 CFU/g 9 .According to Indonesian National Standard Number 7388 of 2009, the maximum microbiological quality requirements for fresh egg food ingredients for Salmonella must be negative per 25 g.
There has been research by Riski L et al. (2023) regarding Salmonella sp in chicken eggs.However, further research must be done regarding Salmonella sp contamination in chicken and duck eggs in other places such as Banjarmasin.So this research aims to analyze Salmonella sp in chicken and duck eggs at the Antasari Traditional Market in Banjarmasin, Indonesia.

MATERIALS AND METHODS
The samples in this study were chicken eggs and duck eggs sold at the Antasari Banjarmasin Traditional Market using a purposive sampling technique with the criteria for cracked/broken eggs and intact eggs, namely three chicken eggs and three duck eggs at six traders so that the total sample was 24.This research was conducted in March 2021.
The tools used in making media are an oven, incubator, hot plate, microscope, analytical balance, knife, blender, petri dish, beaker glass, spoon, stirrer, measuring cup, measuring pipette, erlenmeyer, object glass, colouring rack, spray bottle, spirit lamp, tube, test tube, Durham tube, and dropper pipette.
Informed consent was given to research respondents, namely sellers of chicken and duck eggs at the Antasari Banjarmasin traditional market for availability as research respondents.All procedures of this research comply with the ethical standards of the Declaration of Helsinki.
Egg samples were taken to the laboratory and examined using an ice box.The eggs were removed from the shell aseptically and homogenized using a blender, then 25 ml was taken using a measuring pipette aseptically and planted in Selenite Broth media, incubated for 24 hours at 37 o C. The growth results in Selenite Broth were planted on Salmonella Shigella agar by streaking the loop in a zigzag manner over the surface of the media, then incubated for 24 hours at 37 o C. The suspected colony of Salmonella sp was taken from the Salmonella Shigella agar medium, then made into a Gram stain and planted in sugar media (Glucose, Lactose, Mannitol, Maltose, Saccharose), AP (Peptone Water), VP (Voges Proskauer), MR (Methyl Red), SIM and Simmon Citrate 9 .

RESULTS AND DISCUSSION
Based on the findings of Salmonella sp in chicken and duck eggs sold at the Antasari Banjarmasin traditional market, the results were obtained in Table 1.
Based on Figure 1 Based on the data in Table 1, Salmonella sp was found in 3 samples and other bacteria, namely Pseudomonas sp, Proteus sp, and Klebsiella sp, in 21 samples.The percentage of Salmonella sp identification results can be seen in Figure 1.Based on observations of 6 egg traders, five traders (83.3%) use tables when selling, and one trader (16.7%) does not use a table when selling.The condition of eggs sold as clean (washed first) was not found, and all traders (100%) sold eggs in dirty conditions.The results of observations were that the placement of intact (not broken/not cracked) eggs was separate from broken/cracked eggs in 1 trader (16.7%) and five traders (83.3%) who mixed the placement of the eggs.Observation results showed that one trader (16.7%) had sufficient lighting when selling, and five traders (83.3%) did not have enough lighting when selling.Observation results showed that one trader (16.7%) had a closed trash can, and five traders (83.3%) did not have a closed trash can.The results of observations were that five traders (83.3%) stored eggs in clean and odourless containers and one trader (16.7%) in containers that were unclean and odourless.
Salmonella sp is a pathogenic bacteria that can cause food poisoning.This research was conducted to determine whether or not Salmonella sp was present in chicken eggs and duck eggs sold at the Antasari Banjarmasin traditional market.Based on the results of laboratory examinations, it was found that 12.5% of duck eggs were contaminated with Salmonella sp bacteria.In contrast, none of the chicken eggs were contaminated with Salmonella sp bacteria.In chicken and duck eggs, other bacteria were found, namely Klebsiella sp, as much as 37.5%; Proteus sp, 12.5%; and Pseudomonas sp, 37.5%.The presence of pathogenic bacteria Salmonella sp in food, in this case, the egg samples do not comply with Indonesian National Standard 7388:2009 concerning the maximum limit of microbial contamination in fresh eggs; for Salmonella sp, it must be negative or must not contain Salmonella sp.
Based on observations, it was found that 100% of eggs sold needed to be cleaned first.There are two possible routes for Salmonella contamination of eggs.First, horizontally, the bacteria penetrate the eggshell; secondly, through vertical (transovarian) transmission, the egg's contents are directly contaminated with Salmonella in the reproductive organs before the egg is covered by shell components 11,12 .
The results of this research are in line with the results of research conducted by Loisa et al. (2016), namely that 52 samples of duck meat originating from duck farms in Bogor Regency were tested for the presence of Salmonella sp. with the results of three samples suspected to be positive for Salmonella sp.(5.8%) 13 .
Positive results for Salmonella sp were also obtained in samples of duck eggs that were not intact (broken); eggs that were sold cracked or broken were more easily contaminated with microorganisms around the eggs.The quality of the egg will be better if the eggshell is intact and not cracked.The condition of the eggshell with a rough, cracked and dirty surface will affect the quality of the egg because the eggshell has pores that allow air and dirt to enter the egg.Apart from that, it could also be due to the poor sanitation conditions of the sellers and the market environment that the current condition of the Antasari Banjarmasin traditional market is still simple and does not meet standards.This is proven by the results of observations, which found that the egg storage area was unclean and smelly and did not have a rubbish dump.Which are closed and whole (not broken/not cracked) eggs mixed with eggs that are not whole (broken/cracked).

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According to Cha et al. (2013), the prevalence of Salmonella was 65.2% in duck farms in Pekin, South Korea, with the identification of Salmonella enterica serotypes consisting of S. Typhimurium, S. Enteritidis, and S. London 14 .According to Bouzidi et al.  (2012), Salmonella contamination occurred in laying hen farms, namely S. Enteritidis in Annaba and Eltarf, Algeria 15 .The primary habitat of S. enteritidis is in the digestive tract of warm-blooded animals 16 .S. enteritidis passes through the transovarian contamination route more likely than eggshell penetration 11 .
In the other 21 samples, there were other bacteria, namely Klebsiella sp as much as 37.5%, Proteus sp as much as 12.5%, and Pseudomonas sp as much as 37.5%.According to Morais et al. (2010), generally, the bacteria that contaminate eggs are the Enterobacteriaceae group, such as Shigella sp, Enterobacter sp, Klebsiella sp, Escherichia coli, Proteus sp, and Salmonella sp 17 .
Many other factors may contribute to outbreaks of salmonellosis associated with eggs and egg products 18,19 .Salmonellosis is transmitted through eggs from various dishes such as dressings, pasta, meat/fish pies, custards, and desserts because the food is not heated thoroughly 20 .Salmonella levels in whole eggs are usually less than 10 CFU/egg 21 , but eggs containing more than 105 CFU/g have also been found 21 .Egg whites more often test positive for Salmonella than egg yolks 21 .When stored at room temperature, Salmonella in egg yolks can produce high numbers of Salmonella in a relatively short time 22 .Lowering the temperature to 8°C or lower inhibits the growth of Salmonella in egg yolk 23 .Most previous studies have been conducted at constant temperatures, but there is strong evidence that temperature fluctuations (25 to 35 °C) will increase Salmonella levels in eggs 24 .Storing eggs in the refrigerator from the time of collection and proper cooking will significantly reduce the risk of salmonellosis 25 .A limitation of the research is the small number of samples used, besides manually identifying bacteria using agar media for biochemical tests and checking the test results with references.

CONCLUSION
Eggs at the Antasari Banjarmasin traditional market were contaminated with Salmonella sp as many as 3 (12.5%) of the duck egg samples.Suggestions for consumers are to pay more attention to the hygiene and sanitation of traders before buying eggs, choose clean eggs, store eggs in the refrigerator, and cook eggs until they are cooked.

Figure 1 .
Figure 1.Graph of Percentage of Research Results

Table 1 .
, 12.50% Salmonella sp bacteria were found in 3 samples and 37.50% Klebsiella sp, 12.50% Proteus sp, and 37.50% Pseudomonas sp in 21 other samples.Results of Salmonella sp Examination on Chicken Eggs and Duck Eggs at the Antasari Banjarmasin Traditional Market