Salmonella Serovars Isolated from Poultry Breeding Flocks under the Brazilian Official Control Programme Between 2016 and 2018

ABSTRACT

The control of Salmonella in the poultry production chain combined with biosecurity measures is an important tool to maintain and guarantee the sanitary status of Brazilian flocks. The aim of this work was to compare official laboratory data on molecular typification of Salmonella isolates from poultry breeding flocks in different Brazilian states between 2016 and 2018 and identify the production category with the most positive flocks, in light of current legislation. Surveillance data of positive samples from the official Brazilian Salmonella Control Programme sent to Federal Agricultural Defence Laboratory of São Paulo (LFDA-SP) after molecular characterization were analysed. These data were subject to an exploratory study, undergoing a descriptive statistical analysis followed by the use of frequency and non-parametric hypothesis tests. Overall, 49 serovars were detected in poultry broiler-breeder and layer-breeder flocks. Salmonella ser. Heidelberg, Salmonella ser. Anatum, Salmonella ser. Newport, Salmonella ser. Schwarzengrund and Salmonella ser. Mbandaka were the five most common isolated serovars. The data shows that there is an opportunity to improve biosecurity measures in parent breeder flocks. A total of 16 serovars were identified in turkey-breeders. Salmonella ser. Anatum, Salmonella ser. Newport, Salmonella ser. Brandenburg, Salmonella ser. Litchfield, and Salmonella ser. Livingstone were the most common ones. The four official controlled serovars represented a small part of the isolated strains. These data demonstrate the importance of an official program in Brazil for Salmonella surveillance in breeder flocks combined with biosecurity measures.

Keywords:
Biosecurity; Salmonella official control; Salmonella serovars; Poultry breeders´ categories

INTRODUCTION

Brazil is one of the world’s largest producers of animal protein, the third largest producer of broiler chicken, and the first global exporter of chicken meat and its products. Its privileged position is the result of the country’s natural capacity for producing food, as well as the sanitary status of its flocks. Such position allows industries to stand out in their competitiveness up to the present days (ABPA, 2021).

The southern region of the country, comprising the states of Paraná, Santa Catarina, and Rio Grande do Sul, accounts for approximately 65% of Brazilian chicken meat production and 80% of its exports. On the other hand, the south-eastern region is responsible for the core of the country’s egg production, which has been developing over the years.

Meanwhile, Brazilian turkey meat production has declined considerably after 2017, along with its exports. The southern states of Santa Catarina and Rio Grande do Sul are responsible for 99% of turkey meat exports (ABPA, 2021).

Due to the importance of poultry production, the Ministry of Agriculture, Livestock, and Food Supply (MAPA) launched the National Poultry Health Program (PNSA) in 1994 (Brasil,1994), which has allowed Brazil’s poultry production to move forward. In order to maintain and guarantee the sanitary status of Brazilian flocks, biosecurity and surveillance public policies have been established (Brasil, 2003; 2007; 2016). Nevertheless, the prevention and control of pathogens in those flocks are a permanent challenge, in which the official veterinary service plays a crucial role.

Among such pathogens, Salmonella is a major concern in the poultry production chain worldwide, considering its importance for public and animal health (WOAH, 2021a; WOAH, 2021b). From farm to fork, there are many possible routes for Salmonella dissemination and contamination in poultry and their products, thus examination of each step of the process is necessary (Rajan et al., 2016). Nevertheless, Salmonella is a sanitary barrier to the export of multiplication material (WOAH, 2021b) and poultry products, according to bilateral sanitary agreements in force. Furthermore, the World Organisation for Animal Health (WOAH) suggests that countries should identify the prevalent Salmonella serotypes in humans and poultry in order to establish a control programme (WOAH, 2021a). Therefore, the PNSA implemented the official Salmonella Control Programme (Brasil, 2003).

In this context, the control of this pathogen must start in breeding establishments, considering their potential for dissemination to the rest of the poultry production chain (Sivaramalingam et al., 2013; Rajan et al., 2016). The Brazilian breeding flocks are constantly monitored to be certified as being free from typhoidal and non-typhoidal Salmonella. To monitor Salmonella in the poultry production, samples are collected on a regular basis from all breeder flocks, which includes broiler-breeders, layer-breeders, turkey-breeders, and other bird breeders such as quail, duck, and geese (Brasil, 2003). Additionally, control and monitoring of Salmonella in commercial broilers and turkey establishments was established, as well as measures to control this pathogen in slaughterhouses (Brasill, 2016).

At present, breeding flocks are certified free from 4 (four) Salmonella serovars (Brasil, 2003): Salmonella ser. Enteritidis and Salmonella ser. Typhimurium, which are non typhoidal serovars and important foodborne pathogens worldwide related to poultry products (WOAH, 2021a), and Salmonella ser. Gallinarum e Salmonella ser. Pullorum, typhoidal serovars which concern poultry health and can cause severe economic losses (Markos & Abdela, 2016; Celis-Estupiñan et al., 2017; De Carli et al., 2017; WOAH, 2021b).

The aim of this work was to compare official laboratory data on molecular typification of Salmonella isolates from poultry breeding flocks from different Brazilian states dated between 2016 and 2018, received by the Federal Agricultural Defence Laboratory of São Paulo (LFDA-SP), and to identify the production category with the most positive flocks in light of the current legislation.

MATERIALS AND METHODS

The study analysed the surveillance data of Brazil’s official Salmonella Control Programme, studying positive samples sent to Federal Agricultural Defence Laboratory of São Paulo (LFDA-SP) after molecular characterization. The samples were collected from poultry breeder flocks for certification of epidemiological units, which mostly comprises broiler-breeders, layer-breeders, and a few samples from turkey-breeders, from different states of Brazil (BRASIL, 2003). Other poultry breeder species such as quail-breeders and duck-breeders were excluded from the analyses because of the small number of samples. The data comprises 318 positive Salmonella samples collected in 2016, 2017, and 2018, which are used to describe the major isolated serovars and determine which breeder category had the greatest number of serovars identified.

Types of samples collected in epidemiological units

The surveillance had been performed in broiler-breeders, layer-breeders, and turkey-breeders during rearing and production period in great-grandparent, grandparent, and parent flocks.

Samples were collected between one and five days of life, in the middle of the growing period, at the beginning of production, and then every three months for periodic control of the epidemiological unit.

The type of material used for bacteriological or molecular diagnosis depends on the production stage. In the initial growing phase, dead chicks, drag swabs, and paper from the chick’s transport boxes were collected. Flocks that were vaccinated against paratific Salmonella had their samples collected before vaccination. Cloacal swabs, fresh faeces, and boot swabs were the samples collected during the rearing period from great-grandmothers, grandmothers, and parents. In addition to these, the samples collected in breeders’ flocks vaccinated with inactivated vaccines for paratific Salmonella were pipped embryo, meconium, and organs. After the first certification of the epidemiological unit by the government, the control is conducted on a quarterly basis, collecting samples of cloacal swab, fresh faeces, drag and boot swabs, and pipped embryo (Brasil, 2003).

Salmonella isolation

The samples were sent to accredited laboratories in the National Accredited Laboratories Network. All isolates tested positive for Salmonella in accredited laboratories were sent to the official MAPA laboratory, the Federal Agricultural Defence Laboratory of São Paulo (LFDA-SP), where they are received, identified, purified, and typified by DNA microarray.

Strains purification and biochemical tests

As a confirmation of a diagnosis already made by accredited laboratories with previously isolated strains, the next step performed was purification in agar plate (Rambach). After purification, the colonies were subjected to molecular characterization by DNA microarray test.

Typing

Molecular characterization followed the methodo-logy described in the Routine Salmonella Serotype Identification Check & Trace manual (CHECK-POINTS, 2020). Soon after, three to five colonies isolated on agar plate (Rambach) were collected to be typified when there was suspicion of Salmonella ser. Gallinarum or Salmonella ser. Pullorum, or a single colony for the other Salmonellae. The colony was homogenised in 100 µl of lysis buffer. From the 1.5 mL microtubes, the homogenate was transferred to the thermoblock previously heated to 99°C for 15 min. Afterwards, the homogenate was cooled to room temperature by vortexing. Then, three steps of DNA recognition and detection were performed, as described in the manual. The reading of Salmonella typification results was performed using software available in the analysis system.

Results analysis

Accredited laboratories provided the official laboratory data sheets collection and the Salmonella isolates on a monthly basis. To select the strains considered in this work, the following filters were applied from the database: year, state, region, type of poultry (chicken, turkey, quail and duck); category (e.g. great-grandmother, grandmother); surveillance type (certified as free or controlled epidemiological unit for Salmonella) and type of sample. Exploratory analysis was used on these data, followed by descriptive statistical analysis. Frequency and non-parametric tests (Kruskal-Wallis) were performed using RStudio software.

RESULTS AND DISCUSSION

Surveillance for Salmonella is not only important for public health, but also for the economic losses due to high morbidity and mortality in poultry flocks. Furthermore, Salmonella is responsible for losses and waste of animal protein that reduce global food availability in a world where hunger is a reality. Therefore, Salmonella control in early phases of production could help reducing these losses (FAO, 2011).

Many studies report that Salmonella control and prevention strategies in the poultry chain should cover the whole production chain, from farm to fork (Andino & Hanning, 2015; Rajan et al., 2016; Machado et al., 2020;). Sivaramalingam et al (2013) suggest that interventions at the breeder flock level would likely reduce transmission from breeder flocks to lower levels of the production chain and possibly also to the retail level. When a Salmonella Control Programme for birds is applied at all production stages, including hatcheries, it is possible to keep Salmonella prevalence at a low level (EFSA, 2019). Likewise, the European Commission and European Food Safety Authority (EFSA) attributed the reduction in reported human salmonellosis cases across the European Union to, at least in part, successful control of Salmonella in breeding hens, broiler, laying flocks, and eggs through the National Control Programmes for Salmonella implemented by the United Kingdom in the poultry sector (O’Brien, 2013). Moreover, the National Poultry Improvement Plan (NPIP) executed by the United States Federal Government eradicated Salmonella ser. Pullorum and Salmonella ser. Gallinarum from poultry flocks (Andino & Hanning, 2015). As other countries, in order to reduce Salmonella in the poultry production chain, the Brazilian government implemented a National Surveillance Program for Salmonella in breeder flocks (Brasil, 2003).

The present study analysed data from the National Surveillance Program for Salmonella and the result (Table 1) has shown a great variety of serovars isolated from poultry broiler-breeder and layer-breeder flocks in the main producing states. Some isolates could not be characterised and were described as Salmonella spp., while others were characterised as two possible serovars due to limitations of the microarray assay (CHECK-POINTS, 2020).

Table 1
Salmonella serovars isolated per state from poultry broiler-breeder and layer-breeder flocks, through the Brazilian National Surveillance Programme between 2016 and 2018.

Region	State°		Serovar/year
		2016	2017	2018
	DF	Glostrup	NI*	NI*
Midwest	GO	Enteritidis	Mbandaka	Javiana
		Havana	Ohio	Mbandaka
		Heidelberg	Schwarzengrund	Montevideo
		Infantis		Saintpaul
		Muenster or Reading
		Newport
		Schwarzengrund
	MS	Coeln	Braenderup	Salmonella subspecie non-enterica
		Minnesota	Livingstone
		Schwarzengrund	Newport
			Saintpaul
			Tennessee
	MT	Alachua	Corvallis	Saintpaul
		Corvallis	Salmonella subspecie non-enterica
		Idikan	Saintpaul
		Infantis	Tennessee
		Livingstone
		Meleagridis
		Miami
		Morehead
		Saintpaul
		Salmonella subspecie non-enterica
		Schwarzengrund
North	RO	NI*	Javiana	NI*
	RR	Anatum	NI*	NI*
	TO	Miami	NI*	NI*
Northeast	BA	Miami	Anatum	NI*
	MA	NI*	NI*	Saintpaul
	PB	Muenster or Reading Senftenberg	Sandiego Senftenberg	NI*
	PE	NI*	Salmonella spp.	NI*
South	PR	Anatum	Anatum	Agona
		Enteritidis	Coeln	Anatum
		Give	Corvallis	Braenderup
		Heidelberg	Give	Coeln
		Mbandaka	Heidelberg	Corvallis
		Rissen	Infantis	Gallinarum
		Salmonella 4,[5],12:d:-	Javiana	Give
		Schwarzengrund	Newport	Heidelberg
		Senftenberg	Ohio	Infantis
			Panama	Javiana
			Saintpaul	Mbandaka
			Salmonella 1, 4, [5],12:i:-	Molade
			Schwarzengrund	Muenchen
			Senftenberg	Newport
			Typhimurium	Panama
				Rissen
				Saintpaul
				Schwarzengrund
				Senftenberg
				Thompson Typhimurium
	RS	Javiana	Give	Montevideo
		Minnesota	Mbandaka	Newport
		Montevideo	Saintpaul	Panama
		Senftenberg	Salmonella spp.	Stanley
		Tennessee	Schwarzengrund
		Worthington	Senftenberg
			Stanley
	SC	Anatum	Anatum	Agona
		Genovar 14975	Braenderup	Brandenburg
		Genovar 15101	Brandenburg	Gallinarum
		Gloucester	Give	Heidelberg
		Javiana	Heidelberg	Litchfield
		Livingstone	Idikan	Pullorum
		Mbandaka	Livingstone	Salmonella spp.
		Minnesota	Panama	Senftenberg
			Salmonella spp.
Southeast	MG	Corvallis	Give	NI*
		Panama
		Sandiego
		Schwarzengrund
	RJ	Senftenberg	NI*	NI*
	SP	Anatum	Gloucester	Anatum
		Gallinarum	Livingstone	Gallinarum
		Heidelberg	Minnesota	Gloucester
		Javiana	Senftenberg	Havana
		Minnesota	Typhimurium	Heidelberg
		Schwarzengrund		Isangi
				Livingstone
				Mbandaka
				Minnesota
				Rissen
				Sandiego
				Senftenberg

• *NI: not isolated.
• DF: Distrito Federal; GO; Goiás; MS: Mato Grosso do Sul; MT: Mato Grosso; RO: Rondônia; RR; Roraima; TO: Tocantins, BA: Bahia; MA: Maranhão; PB: Paraíba; PE: Pernambuco; PR: Paraná; RS: Rio Grande do Sul; SC: Santa Catarina; MG: Minas Gerais; RJ: Rio de Janeiro; SP: São Paulo.

Overall, 49 different serovars were detected in poultry broiler-breeder and layer-breeder flocks throughout the three analysed years. Salmonella ser. Heidelberg (10,7%), Salmonella ser. Anatum (6,3%), Salmonella ser. Newport (6,3%), Salmonella ser. Schwarzengrund (5,9%) and Salmonella ser. Mbandaka (5,1%) were the five most common isolated in poultry breeder flocks, accounting for 34,4% of all characterised strains (Figure 1). According to our findings, a Canadian study based on their official data on those types of breeder flocks, between 1998 and 2008, identified the majority of the serovars isolated in Brazil (Sivaramalingam et al., 2013). Salmonella ser. Heidelberg was the most common serovar in both studies.

Figure 1
Fifthteen most common serovars isolated in poultry broiler-breeder and layer-breeder flocks between 2016 and 2018.

In terms of Brazilian regions, 39 distinct serovars were identified in the Southern region, where most breeders’ establishments are located. In the Midwestern region, 25 distinct serovars were identified, while in the southeast region there were 18 distinct serovars. On the other hand, only four and six distinct serovars were isolated through the analysed years in the Northern and Northeastern regions, respectively (Table 1). The most common isolated serovars in the Southern region were Salmonella ser. Heidelberg, Salmonella ser. Newport, and Salmonella ser. Anatum. Salmonella breeder data in Brazil are scarce and a great number of studies are based on broiler data. Some of those studies have demonstrated the presence of Salmonella ser. Heidelberg in broiler establishments in the Southern region (Pandini et al., 2015; Voss-Rech et al., 2015) which emphasises that this serovar is more resistant than others in the environment (Voss-Rech et al., 2019). Indeed, our study shows that Salmonella ser. Heidelberg has been constantly isolated in this region, especially in the two main producer states, Paraná and Santa Catarina (Figure 2).

Figure 2
Most common serovars isolated in the Southern region of Brazil in poultry broiler-breeder and layer-breeder flocks between 2016 and 2018.

Our results demonstrated that the number of serovars identified in Paraná and São Paulo has increased through the analysed years (Table 1). A review was performed on the number of poultry establishments through those years and the increase in the variety of serovars could not be related to the increase in the number of poultry producing establishments. On the other hand, layer breeder population and egg production have clearly risen in those years, which might have contributed for the increase of Salmonella isolations in breeder flocks; particularly in São Paulo, where the production of laying hens is predominant (ABPA, 2021). Another possibility to explain the increase in the number of serovars detected is a failure in the biosecurity programs, which may facilitate the spread of local Salmonella strains (De Carli et al., 2017).

Some serovars are common in broiler-breeders regardless of the production region. Data analysis has shown that the frequency of appearance of different serovars in poultry production in the regions of Brazil is statistically significant. The Southern and Midwestern regions presented the greatest variety of serovars throughout the analysed period.

Figure 3
Frequency of distinct serovar in Brazil per region in poultry broiler-breeder and layer-breeder flocks between 2016 and 2018.

The aim of Brazilian breeding flock’s Salmonella certification is to eradicate the four official controlled serovars from great-grandparent and grandparent flocks; to eradicate Salmonella ser. Pullorum and Salmonella ser. Gallinarum from parent flocks; and to control Salmonella ser. Enteritidis and Salmonella ser. Typhimurium in parent flocks (Brasil, 2003). In addition to the Salmonella surveillance programme, poultry breeder establishments have also implemented compulsory biosecurity measures in order to prevent and control pathogens (Brasil, 2007).

The four serovars controlled by the official surveillance programme, Salmonella ser. Enteritidis, Salmonella ser. Typhimurium, Salmonella ser. Gallinarum and Salmonella ser. Pullorum, represent a small part of the strains isolated (5,5%). Among those serovars, Salmonella ser. Gallinarum was the most common one isolated, followed by Salmonella ser. Typhimurium, Salmonella ser. Enteritidis, and Salmonella ser. Pullorum (Figure 4). The results show that the Salmonella control programme combined with biosecurity measures (Brasil, 2007) is fulfilling its purpose, even though the aim of the official control program has not been achieved.

Figure 4
Percentage of controlled serovars isolated in Brazil in poultry broiler-breeder and layer-breeder flocks between 2016 and 2018.

Regarding the four serovars, a Canadian survey showed that less than 1% of Salmonella-positive isolates in breeders between 1998 and 2008 were Salmonella ser. Enteritidis, whereas Salmonella ser. Gallinarum and Salmonella ser. Pullorum were not isolated. Although in the same study Salmonella ser. Typhimurium was the fourth most common serovar, it represented only 2.8% of the isolates (Sivaramalingam et al., 2013). Costa et al. (2013) suggested that the decline in the prevalence of Salmonella ser. Enteritidis and the lack of identification of Salmonella ser. Pullorum and Salmonella ser. Gallinarum in commercial poultry carcass and poultry products in Brazil might reflect the actions of the government programs geared to its control. Nevertheless, those serovars can still be isolated, which may indicate biosecurity gaps.

Salmonella phylogenetic analysis shows common populations and similar strains of Salmonella ser. Enteritidis between humans and chickens, considering that poultry products are the primary vehicle of this serovar transmission to humans, this may be attributable to infected breeding flocks (Li et al., 2021). Moreover, the control of Salmonella ser. Gallinarum and Salmonella ser. Pullorum can be complicated due to vertical transmission, as hens can become sub clinically infected carriers and pass the infection on to their embryos (Berhanu & Fulasa, 2020). Consequently, a lot of effort still needs to be made to eradicate and control those serovars in the poultry chain.

Poultry breeder categories differed in the number of positive epidemiological units for Salmonella. The percentage of positive Salmonella epidemiological units is much higher in parents than in other categories such as great-grandparent and grandparent poultry in all three years. Even in the rearing period, parent-breeder was the category with the most Salmonella positive epidemiological units identified, despite the fact that the same legal biosecurity measures are required for all breeder categories (Table 2).

Table 2
Percentage of Salmonella positive epidemiological unit per broiler-breeder and layer-breeder category and year in Brazil.

Breeder category	(%) Positive epidemiological units /year
	2016	2017	2018
Great-grandparent - rearing period	0	0	0
Great-grandparent	1	0	0
Grandparent - rearing period	0	0	1
Grandparent	9	0	1
Parent - rearing period	0	11	15
Parent	90	89	83

Current knowledge about risk/protective factor categories in relation to the farming system of breeder flocks to improve Salmonellae control highlight outdoor access, group size, stocking density, genetic, farm hatching, existence of enrichment (such as perches, windows, nest boxes), type of litter, factors relating to biosecurity measures (e.g. disinfection, pest control), and occurrence of other diseases. Also, animal welfare indicators listed were stress (other than heat stress), heat stress, activity/behaviour, body status (e.g. foot pad dermatitis - FPD) and other diseases (EFSA et al., 2019).

Although biosecurity legal requirements are the same for these different breeder categories (Brasil, 2007), great-grandparents flocks probably have a higher sanitary status, which implies more strict biosecurity measures. Based on these study data, it is possible to assume that the correct application of biosecurity measures may differ between categories. Therefore, data shows that there is an opportunity to improve biosecurity measures in parent breeder flocks.

When the data about turkey breeder flocks was analysed, a total of 16 serovars were identified. The most common isolated serovars were Salmonella ser. Anatum, Salmonella ser. Newport, Salmonella ser. Brandenburg, Salmonella ser. Litchfield, and Salmonella ser. Livingstone, representing 72% of identified strains. It is important to highlight that none of the official controlled serovars were detected (Table 3).

Table 3
Salmonella serovars isolated per state from poultry turkey-breeder flocks, through the Brazilian National Surveillance Programme between 2016 and 2018.

Year	Breeder category	State/Serovar
		GO	MG	SP	RS	SC
2016	Great-grandparent	NI*	NI*	NI*	NI*	Newport
		Anatum	Anatum	NI*	Anatum	NI*
		Brandenburg	Genovar 15083		Brandenburg
	Parent	Genovar 14973 (poss. Javiana)	Newport		Genovar 6255
		Genovar 15101			Livingstone
		Heidelberg			Senftenberg
		Schwarzengrund
2017		NI*	NI*	NI*	Brandenburg	Anatum
	Parent				Give
					Senftenberg
2018	Parent - rearing period	NI*	NI*	NI*	Corvallis
		NI*	NI*	NI*	Corvallis	Agona
	Parents				Senftenberg	Anatum
					Tennessee	Litchfield
						Newport

• *NI: not isolated.

The data presented on Table 3 differs from other studies which isolated Salmonella ser. Typhimurium and Salmonella ser. Enteritidis in turkey facilities, even though with very low prevalence (O´Brian, 2013; Sivaramalingam et al., 2013). Another research regarding official data with Salmonella isolates from turkey carcasses was realised in the Southern region of Brazil, between 2004 and 2006, and identified the occurrence of Salmonella ser. Enteritidis and Salmonella ser. Typhimurium in turkey products (Palmeira et al., 2016). Brazil’s turkey production is currently represented by a few establishments. The lack of isolation of official serovars in this study may be caused by the ongoing official certification focused on the eradication and control of those serovars and the decrease in Brazilian turkey production (ABPA, 2021).

We analysed different types of samples used to certify the breeder flocks. It was possible to asses that most Salmonella positive isolates were from drag swabs, boot swabs, and pipped embryo samples (Table 4).

Table 4
Type of samples collected and corresponded Salmonella serovars isolated by year in broiler-breeder and layer-breeder in Brazil.

Type of sample	Sampling year/serovar
	2016	2017	2018
Boot swabs	Anatum	Anatum	Agona
	Enteritidis	Give	Anatum
	Genovar 14975	Gloucester	Corvallis
	Gloucester	Heidelberg	Give
	Heidelberg	Idikan	Gloucester
	Idikan	Infantis	Heidelberg
	Infantis	Minnesota	Isangi
	Javiana	Newport	Javiana
	Livingstone	Panama	Litchfield
	Minnesota	Saintpaul	Livingstone
	Muenster or Reading	Schwarzengrund	Mbandaka
	Newport	Senftenberg	Minnesota
	Saintpaul	Stanley	Montevideo
	Schwarzengrund	Typhimurium	Muenchen
	Senftenberg		Newport
			Saintpaul
			Senftenberg
			Stanley
Chick box liners	Minnesota	NI*	Livingstone
	Senftenberg		Newport
			Rissen
			Sandiego
			Salmonella subspecie non-enterica
Cloacal swabs	Miami	Sandiego	NI*
	Panama	Senftenberg
	Schwarzengrund
	Senftenberg
Pipped embryo	Alachua	Braenderup	Agona
	Corvallis	Corvallis	Anatum
	Gallinarum	Newport	Gallinarum
	Infantis	Ohio	Heidelberg
	Livingstone	Panama	Mbandaka
	Mbandaka	Salmonella spp.	Newport
	Meleagridis	Senftenberg	Panama
	Miami		Schwarzengrund
	Minnesota		Senftenberg
	Schwarzengrund		Thompson
Dead day-old chicks	Glostrup	Heidelberg	Montevideo
	Genovar 15101	Livingstone	Newport
	Senftenberg	Typhimurium
Drag swabs	Anatum	Anatum	Agona
	Enteritidis	Braenderup	Anatum
	Give	Brandenburg	Braenderup
	Havana	Coeln	Coeln
	Heildelberg	Corvallis	Give
	Miami	Give	Havana
	Montevideo	Heidelberg	Heidelberg
	Morehead	Javiana	Infantis
	Salmonella 4,[5],12:d:-	Livingstone	Javiana
	Salmonella subspecie non-enterica	Mbandaka	Mbandaka
	Sandiego	Newport	Molade
	Schwarzengrund	Ohio	Newport
	Tennessee	Panama	Panama
		Saintpaul	Saintpaul
		Salmonella subspecie non-enterica	Sandiego
		Salmonella spp.	Schwarzengrund
		Schwarzengrund	Senftenberg
		Tennessee	Typhimurium
Faecal	Coeln	Anatum	NI*
	Miami	Heidelberg
Meconium	Mbandaka	Anatum	Livingstone
	Meleagridis	Corvallis	Senftenberg
	Muenster or Reading	Give
	Rissen	Mbandaka
	Senftenberg	Tennessee
		Typhimurium
Organs	Corvallis	Saintpaul	Brandenburg
	Salmonella subspecie non-enterica	Salmonella 1, 4, [5], 12:i:-	Gallinarum
			Heidelberg
			Pullorum
			Salmonella spp.
Litter swab	Javiana	Senftenberg	NI*
	Worthington

• *NI: not isolated.

Regarding different sample types used to isolate Salmonella in breeders, Berghaus et al. (2012) conducted a serial survey of 49 broiler breeder farms across the United States of America. Different sample types such as drag swabs, boot swabs, and litter samples yielded comparable prevalence of Salmonella positive results, while the prevalence was significantly lower for the slat and egg belt sponge samples. The study by Kahya et al. (2013) compared different types of samples for Salmonella detection in chicken layer breeder flock, and positive Salmonella samples were 8% out of pooled cloacal swab, 90,9% out of pooled embryonated chicken egg, and 21,4% of pooled wet faeces. PCR and ISO 6579 culture methods results were 100% in agreement (100% sensitivity and specificity) with culture results for all sample types. These results showed that pipped embryo was the most meaningful sample for Salmonella detection from breeder flocks, followed by wet faeces.

As expected, Salmonella ser. Gallinarum and Salmonella ser Pullorum were isolated from organs. According to the WOAH (2021b), post mortem tissues are preferable to isolate those serovars. It is important to highlight Salmonella ser. Gallinarum isolationfrom dead embryo. This matrix is easy to collect and include in poultry industries’ biosecurity programs. Finally, Salmonella ser. Enteritidis, was detected from boot and drag swabs and Salmonella ser. Typhimurium was isolated from a large number of sample sources, like day old chicks, meconium, boot, and drag swabs. These data reinforce the importance of collecting tissues to detect typhoid Salmonella and suggest that the embryo has a great value in monitoring the presence of this pathogen.

CONCLUSION

Salmonellosis is a disease of economic and public health importance throughout the world, especially for those countries that are major exporters of poultry products, such as Brazil, requiring constant monitoring of the prevalent strains and their characteristics. Especially in the case of Brazil, the adoption of the verticalized poultry production model requires special attention in relation to the control of poultry salmonellosis in breeder flocks.

Our results showed that the number of serovars has increased between 2016 and 2018 in the main broiler and layer poultry producer states; the frequency of distinct serovars in the different regions of Brazil were statistically significant; parent-breeders was the category with the most Salmonella positive epidemiological units identified; and Salmonella ser. Enteritidis, Salmonella ser. Typhimurium, Salmonella ser. Gallinarum, and Salmonella ser. Pullorum represent a small part of the isolated strains.

Therefore, the present study demonstrates the importance of an official program in Brazil for the surveillance for salmonella in breeder flocks, especially when combined with biosecurity measures designed to prevent and control the disease.

ACKNOWLEDGEMENT

To the Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), ProcessE-26/211.075/2021- Edital E-29-Apoio aos Programas de Pós Graduação Stricto Senso do Estado Do Rio de Janeiro, for financial support for the publication of this work.

REFERENCES

• ABPA - Associação Brasileira de Proteína Animal. Relatório anual ABPA 2021 [cited 2021 Nov 10). Available from: https://abpa-br.org/relatorios/.
  » https://abpa-br.org/relatorios
• Andino A, Hanning I. Salmonella enterica: survival, colonization, and virulence differences among serovars. The Scientific World Journal 2015;2015.
• Berghaus RD, Mathis DL, Bramwell RK, Macklin KS, Wilson JL, Wineland MJ, et al. Multilevel analysis of environmental salmonella prevalences and management practices on 49 broiler breeder farms in four South-Eastern States, USA. Zoonoses and Public Health 2012;59(5):365-74.
• Berhanu G, Fulasa A. Pullorum disease and fowl typhoid in poultry: a review. British Journal of Poultry Sciences 2020;9(3):48-56.
• Brasil. Ministério da Agricultura, Pecuária e do Abastecimento. Portaria ministerial nº193, de 19 de setembro de 1994. Institui o programa nacional de sanidade avícola no âmbito do dsa e cria o comitê consultivo do programa de sanidade avícola. Diário Oficial da União, Seção I; 1994.
• Brasil. Ministério da Agricultura, Pecuária e do Abastecimento. Instrução normativa nº 78, de 03 de novembro de 2003. Normas técnicas para controle e certificação de núcleos e estabelecimentos avícolas como livres de Salmonella Gallinarum e de Salmonella Pullorum e Livres ou Controlados para Salmonella Enteritidis e para Salmonella Typhimurium. Diário Oficial da União, Seção I; 2003.
• Brasil. Ministério da Agricultura, Pecuária e do Abastecimento. Instrução normativa nº 56, de 04 de dezembro de 2007. Procedimentos para registro, fiscalização e controle de estabelecimentos avícolas de reprodução, comerciais e de ensino ou pesquisa. Diário Oficial da União, Seção I; 2007.
• Brasil. Ministério da Agricultura, Pecuária e do Abastecimento. Instrução normativa nº 20, de 21 de outubro de 2016. Estabelece o controle e o monitoramento de Salmonella spp. nos estabelecimentos avícolas comerciais de frangos e perus de corte e nos estabelecimentos de abate de frangos, galinhas, perus de corte e reprodução, registrados no Serviço de Inspeção Federal (SIF). Diário Oficial da União, Seção I; 2016.
• Celis-Estupiñan ALDP, Batista DFA, Cardozo MV, De-Souza AIS, Alves LBR, De-Almeida AM, et al. Further investigations on the epidemiology of fowl typhoid in Brazil. Avian Pathology 2017;46(4):416-25.
• Check-Points. Check & trace Salmonella user manual version 2020-03 [cited 2021 Nov 25). Available from: https://www.woah.org/app/uploads/2021/03/official-cts-user-manual-shortened-version-2020-09.pdf
  » https://www.woah.org/app/uploads/2021/03/official-cts-user-manual-shortened-version-2020-09.pdf
• Costa RG, Festivo ML, Araujo MS, Reis EM, Lázaro NS, Rodrigues DP. Antimicrobial susceptibility and serovars of Salmonella circulating in commercial poultry carcasses and poultry products in Brazil. Journal of Food Protection 2013;76(12):2011-7.
• De Carli S, Gräf T, Kipper K, Lehmann FKM, Zanetti N, Siqueira FM, et al. Molecular and phylogenetic analyses of SalmonellaGallinarum trace the origin and diversification of recent outbreaks of fowl typhoid in poultry farms. Veterinary Microbiology 2017;212:80-6.
• EFSA BP, Koutsoumanis K, Allende A, Alvarez-Ordóñez A, Bolton D, Bover-Cid S, et al. Salmonella control in poultry flocks and its public health impact. EFSA Journal 2019;17(2):e05596.
• FAO - Food and Agriculture Organization of the United Nations. Global food losses and food waste - Extent, causes and prevention; 2011 [cited 2021 Nov 10]. Available from: https://www.fao.org/3/i2697e/i2697e.pdf.
• Kahya S, Eyigör A, Çarli KT. Comparison of different sample types for Salmonella detection from chicken layer breeder flocks. Uludag Üniversitesi Veteriner Fakültesi Dergisi 2013;32(2):19-24.
• Li S, He Y, Mann DA, Deng X. Global spread of Salmonella Enteritidis via centralized sourcing and international trade of poultry breeding stocks. Nature Communications 2021;12(1):1-12.
• Machado SCA, Pereira VLA, Aquino MHC, Giombeli A, Rodrigues DP, Nascimento ER. Qualitative and quantitative analysis of salmonella spp. in broilers technological processing and determination of a performance objective (PO) for frozen chicken breast. Brazilian Journal of Poultry Science 2020;22(1):1-12.
• Markos T, Abdela N. Epidemiology and economic importance of pullorum disease in poultry: A review. Global Veterinaria 2016;17:228-37.
• O'Brien SJ. The "decline and fall" of nontyphoidal Salmonella in the United Kingdom. Clinical Infectious Diseases 2013;56(5):705-10.
• Palmeira A, Santos LR, Borsoi A, Rodrigues LB, Calasans M, Nascimento VPD. Serovars and antimicrobial resistance of Salmonella spp. isolated from turkey and broiler carcasses in southern Brazil between 2004 and 2006. Revista do Instituto de Medicina Tropical de São Paulo 2016;58.
• Pandini JA, Pinto FGDS, Muller JM, Weber LD, Moura ACD. Ocorrência e perfil de resistência antimicrobiana de sorotipos de Salmonella spp. isolados de aviários do Paraná, Brasil. Arquivos do Instituto Biológico 2015;82:1-6.
• Rajan K, Shi Z, Ricke SC. Current aspects of Salmonella contamination in the US poultry production chain and the potential application of risk strategies in understanding emerging hazards. Critical Reviews in Microbiology 2016;43(3):370-92.
• Sivaramalingam T, McEwen SA, Pearl DL, Ojkic D, Guerin MT. A temporal study of Salmonella serovars from environmental samples from poultry breeder flocks in Ontario between 1998 and 2008. Canadian Journal of Veterinary Research 2013;77(1):1-11.
• Voss-Rech D, Vaz CS, Alves L, Coldebella A, Leao JA, Rodrigues DP, et al. A temporal study of Salmonella enterica serotypes from broiler farms in Brazil. Poultry Science 2015;94(3):433-41.
• Voss-Rech D, Kramer B, Silva VS, Rebelatto R, Abreu PG, Coldebella A, et al. Longitudinal study reveals persistent environmental Salmonella Heidelberg in Brazilian broiler farms. Veterinary Microbiology 2019;233:118-23.
• WOAH - World Organisation for Animal Health. Prevention, detection and control of Salmonella in poultry. In: WOAH. Terrestrial animal health code; 2021a [cited 2021 Nov 10]. Available from: https://www.woah.org/en/what-we-do/standards/codes-and-manuals/terrestrial-code-online-access/?id=169&L=1&htmfile=chapitre_prevent_salmonella.htm
  » https://www.woah.org/en/what-we-do/standards/codes-and-manuals/terrestrial-code-online-access/?id=169&L=1&htmfile=chapitre_prevent_salmonella.htm
• WOAH - World Organisation for Animal Health. Fowl typhoid and pullorum disease. In: WOAH. Terrestrial animal health code; 2021b [cited 2021 November 10]. Available from: https://www.woah.org/en/what-we-do/standards/codes-and-manuals/terrestrial-code-online-access/?id=169&L=1&htmfile=chapitre_fowl_typhoid_pullorum_disease.htm
  » https://www.woah.org/en/what-we-do/standards/codes-and-manuals/terrestrial-code-online-access/?id=169&L=1&htmfile=chapitre_fowl_typhoid_pullorum_disease.htm

Publication Dates

• Publication in this collection
  20 Jan 2023
• Date of issue
  2023

History

• Received
  15 Mar 2022
• Accepted
  25 July 2022