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In ternationa l
Scholars
Journa ls

 

African Journal of Pig Farming ISSN: 2375-0731 Vol. 9 (1), pp. 001-006, January, 2021. Available online at 
www.internationalscholarsjournals.org © International Scholars Journals 

 

Author(s) retain the copyright of this article. 

 
 
Full Length Research Paper 

 

Seasonal emergence of swine erysipelas in hilly 
state Nagaland, Northeast India 

 
Nagendra Nath Barman1*, Debojyoti Borkotoky2, Biswajyoti Borah1, Anjan Jyoti 

Nath3, Papiya Das1 and Durlav Prasad Borah1
 

 
1
Department of Microbiology, College of Veterinary Science, Assam Agricultural University, Khanapara campus, 

Guwahati-781022, Assam, India. 
2
Subject Matter Specialist (Animal Science), KVK Phek, ICAR-NRC on Mithun, Nagaland-797007,  India. 

3
Department of Microbiology, Lakhimpur College of Veterinary Science, Assam Agricultural University, Joyhing, North 

Lakhimpur- 787 051, Assam, India. *Corresponding author. E-mail: nnbarman@gmail.com. 
 

Accepted 20 September, 2019  
 

Seasonal outbreaks of swine erysipelas have been reported in back yard pig farms in the Phek district 
of Nagaland, India. The alpha haemolytic isolate of Erysipelothrix rhusiopathiae was recovered on blood 
agar from the clinical samples. The organisms were confirmed microscopically, biochemical analysis as 
well as by polymerase chain reaction (PCR) amplification of 16S rRNA gene and sequence analysis. 
These Nagaland isolates (KT160358, KT160359) were closely related to the type spp. E. rhusiopathiae in 
phylogenetic analysis and forms the same clad with Chineese isolates of swine and murine origin 
indicating an epidemiological link. The isolates were found to be most sensitive to oxytetracycline and 
responded to treatment. Swine erysipelas occurred in Phek district in a season due to sudden change 
of weather and temperature. Pigs exposed to such predisposing factors probably favoured to 
propagation of already persisted organisms in pigs. This is the first confirmed case of E. rhusiopathiae 
infection from the NE states of Nagaland, India. 

 
Key words: Swine erysipelas, Erysipel othri x rhusiopathiae, pig, polymerase chain reaction (PCR) Nagaland, 
India, Oxytetracycline.  

 
 
INTRODUCTION 

 
Erysipelothrix rhusiopathiae, belonging to the family 
Erysipelotrichac eae, is a non-motile, Gram-positive, non-
sporulating, non-acid-fast organism distributed worldwide 
affecting wide variety of vertebrate and invertebrate 
species including man (Reboli and Farrar, 1989). 
Organisms in many occasions harbour by pigs in lymph 
nodes and shed along with feces, urine, saliva and nasal 

 
 
secretions (Lee et al., 2011). Affected pigs manifest the 
disease as (i) acute septic form, (ii) subacute urticarial 
form marked by reddis h-purple rhomboid spots or 
"diamonds" in the skin, (iii) joint or arthritic form, and (iv) 
chronic cardiac form (endocarditis) (Reboli and Farrar, 
1989).Various predisposing factors, change of 
environmental conditions and parasitic infestation lead to  



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reappearance of s wine erysipelothrix (SE) infection in 
that population. Seasonal outbreaks of swine erysipelas 
were investigated in Phek district of Nagaland, India 
during 2013-2015. 

 

MATERIALS AND METHODS 
 
Outbreaks of Sw ine erysipelas – samples collection 
 
The disease of sw ine erysipelas w as reported from Porba village (altitude 

1985 MSL), District Phek of Nagaland during the summer rainy season from 

2013 to 2015. The village had a pig population of 552 cross bred and 200 

local doom pigs as per the livestoc k census report 2012 (GOI, 2012). Far 

mers keep pigs as a bac k yard s mall unit mainly for meat purpose and fed 

them on household as w ell as hotel w aste. In every rainy summer season ( 

May – July) there w as disease outbreaks in pigs. Affected animals (150) 

clinically w ere anorexic and recumbent w ith high fever (106°F) for 2-3 days. 

Some  
animals developed erythematous patches. Postmortem examination 
revealed haemorrhages in intestine, congestion of liver, spleen and 
kidney. During the course of investigation 3 more animals died w 
ithin 12 days. All affected pigs w ere also reported to be infested w 
ith pig louse Haemat opinus suis. Clinical sample like sw abs from 
the w ound and tissue biopsy from the affected areas were 
collected in sterile containers for isolation of the causative organis 
m. Samples also included sloughed off tissues and biopsy samples 
preserved in 10% formalin for histopathological study . 

 

Virological investigation 
 
Tissue samples w ere  processed  for demonstration of classical  
sw ine fever using single step Reverse transcription poly merase 
chain reaction ( RT- PCR) (Hoffmann et al., 2005) and sw ine pox 
as per the method of Medaglia et al. (2011). 

 
Identification and antibiotic activity of Erysipelothrix 
rhusiopathiae 
 
The sw ab samples ( N=51) from the affected areas w ere 
inoculated into nutrient broth and incubated aerobically at 37°C for 
48 h. Sub-culturing w as done on blood agar plates, incubated at 
37˚ C in presence of 5% CO2 for 24 h. Colony mor phologies w ere 
studied. Gram’s staining and biochemical analysis w ere done to 
confirm the organis m. A panel of antibiotic discs containing 
amikacin (30 µg), amoxicillin (30 µg), ampicillin (10 µg), cefotaxime 
(30 µg), chloramphenicol (30 µg), c iprofloxacin (5 µg), cloxac illin 
(30 µg), co-trimoxazole (25 µg), enrofloxacin (5 µg), gentamicin (10 
µg), neomycin (30 µg), norfloxacin (10 µg), oxytetracycline (30 µg), 
streptomycin (25 µg) and tetracycline (30 µg) w as used to study 
the antibiotic sensitivity patter n of the isolates. The zone of 
inhibition was measured, recorded and interpreted according to the 
clinical and Laboratory Standards Institute criteria (CLIS-MIC). 

 
Detection of nucleic acid of Erysipelothrix rhusiopathiae and 
sequencing 
 
For detection of E. rhusiopathiae nucleic acid tissue samples w ere 

processed using DNA Sure® Tissue Mini Kit ( Nucleo-pore, cat.#NP-61305) 

and poly merase chain reaction ( PCR) amplification 
for 16S r RNA gene pr imers ( MO101 F  
AGATGCCA TAGAAACTGGTA and M0102 R CTGTATCCGCCA 
TAA CTA) of E. rhusiopathiae ( Makino et al., 1994) w as used. The 
PCR conditions w ere optimized w ith a final volume of 25 µl at 
94°C for 5 min follow ed by 30 cycle of 94°C for 30 s, 54˚C for 2 
min and 72˚C for 45 s and final extension w as 

 
 
 
 

 
carried out at 72°C for 5 min. In PCR r eaction 28 ng/ µl total genomic DNA 

w as taken along w ith positive and negative control. Pr oducts of PCR w ere 

visualized in 2% agarose gel electrophoresis under Geldoc ( Kodak, USA). 

Further PCR products w ere purified by QIAquick PCR purification kit 

protocol and sequenced. 

 

Analysis of gene sequence and phylogenetic studies on 
Erysipelothrix rhusiopathiae 
 
To deter mine the relationship of the Nagaland isolates of E. rhusiopathiae w 

ith other isolates of this species, 16S r RNA gene was amplified and 

sequenced (GenBank accession number KT160358 and KT160359). 

Sequence identity at nucleotide level was deter mined by Clustal W method 

of Meg-Align program in DNA STA R pac kage ( DNASTA R Inc., USA). The 

phylogenetic tree was constructed using other 16S r RNA gene sequences 

available at NCBI (viz. strain ZYL(KF811052.1), Isolate EU188793.1, 
 
strain:KG- BB1(AB055909.1), strain Fujisaw a(NR_074878.1), 
Isolate  DQ462571.1,  strain: A TCC 19414(AB055905.1), strain 
JPB251209S( HM569359.1), Chiba9393( EF494748.1), sp.  
T127_5( JQ739693.1), sp. LV19(KJ670316.1), strain Er.GXLC-

1(KP063151.1), inopinata strain 143-02 ( inopinata strain 143-02) strain: 

JCM 8534(LC019778.1) and strain Er.GXBY -1(KP063149.1). Sequences w 

ere aligned by ClustalX version 2.1 (www .clustal.org), and the concatenated 

alignments w ere used for phylogeny inference ( MEGA5; www .megasoftw 

are.net) opting for the Maximum parsimony and Poisson correction. 

Computed r eplicates for bootstrap support w as done and values w ere 

observed. 

 

Histopathology 
 
For malin fixed tissues w ere processed as per standard protocol for 
histopathological studies. Sections of 4-5 micron thickness w ere 
stained routinely w ith Haematoxylin and Eos in stain and observed 
under oil immersion objective of a low pow er light microscope. 
 
 

RESULTS AND DISCUSSION 

 

Tissue samples processed for detection of classical 
swine fever virus and swine pox virus were confirmed as 
negative for both viral agents. However, bacteriological  
investigation demonstrated association of E. 
rhusiopathiae infection in affected (5) as well as in dead  
(2) pigs. Prevalence of erysipelothrix in many animals, mostly 

in pigs and birds has been reported throughout the world 

including from India (Shank ar et al., 2009; Arora et al., 2011). 

But there was a single report on swine erysipelothrix (SE) from 

Meghalaya (Das et al., 2014). North Eastern Region has the 

highest pig population of the country. Diverse geographical 

locations, varied climatic situations and frequent movement of 

pigs favour for spread of the disease through carrier pigs (Leslie 

et al., 2015). Present report is a thorough investigation on 

 
swine erysipelothrix occurred seasonally at Nagaland, 
another NE state of India. 
 
 
Clinical signs 

 
A total of 150 pigs during 2013-15 exhibited clinically high 

fever (105-107
0
F), anorexia, firm faeces, and animals 



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Figure 1. Character istic lesions of sloughed off skin from the cases of  E. 
rhusiopathiae infected pigs. 

 
 

 

lying down. Cutaneous lesions appeared after 5-7 days 
of illness initiating with erythematous patches followed by 
papules of 4-5 mm diameter, dark purple, raised, firm to 
touch giving square to rhom boid shape in entire body. 
No vesicular stage was noticed. Although scabs 
developed in the entire body, prominent lesions were 
identified on head, face back, belly, limbs, tail and on ear 
(Figure 1). In unattended cases scab lesions were 
sloughed off within 17-22 days leaving a large ulcerated 
area. Thes e typical cutaneous lesions have been mostly 
seen in grower animals. In affected pigs it appears that 
cutaneous lesions were developed following acute stages 
of illness.  

Considering the pathogenesis of E. rhusiopathiae, 
organisms gain access to the body, probably through the 
tonsils or other lymphoid tissue of the digestive tract and 
spread throughout the body. The bacteria produce  
neuraminidas e, an enzyme that cleaves 
mucopolysaccharides in cell walls which may mediate the 
wides pread vascular damage that accompanies SE. 
Vascular damage leads to thrombosis and interference 
with microcirculation in capillaries and venules at many 
sites. Classic cutaneous rhomboid urticaria (diamond 
skin) occurs in a percentage of pigs shortly after the 
acute febrile stages. In younger pigs with acute 
erysipelas, signs are similar, with cyanosis of extremities, 
ears and snouts pronounced and urticaria less common 
(Anonymous, 2016).  

Seasonal occurrence of swine erysipelas in Nagaland 
justifies the stress due to sudden change of temperature. 
As stated by Amanda (2012)that stress factors such as 
overstocking, mixing pigs after weaning, and sudden 
changes in temperature can trigger clinical erysipelas. 

 
 
 

 

Isolation and identification of organism 

 

On blood agar the organism produced small, circular and 
transparent α- hemolytic colonies with a smooth 
glistening surface and edge. Biochemically all the 
isolates were catalase, oxidase and urease negative, 

produces H2S and ferments glucose and lactose. All 7 

isolates were identified as E. rhusiopathiae based on the 
cultural, morphological and biochemical characteristics. 
The Nagaland isolates were sensitive in vitro to 
oxytetracycline, tetracycline, ampicillin, amoxicillin and 
cloxacillin; moderately sensitive to streptomycin, 
enrofloxacin, amikacin, co-trimoxazole, cefotaxime and 
ciprofloxacin; and resistant to gentamicin and norfloxacin, 
chloramphenicol and neomycin. Based on the 
antibiogram, survived ailing pigs were treated with 
oxytertacycline at 10 mg/kg body weight, intramuscularly 
for one week. Eight out of 13 treated animals responded 
promptly and recovered. Treatment of swine erysipelas 
cases with penicillin (Shankar et al., 2009) or other 
penicillin group of drugs such as the combination of 
amoxycillin and cloxacillin (Das et al., 2014) have been 
frequently reported. 
 

 

Molecular confirmation and characterization 

 

Tissue samples were negative for Swine fever virus and 
pox virus in PCR. A total of seven E. rhusiopathiae 
isolates recovered from tissue samples were subjected 
for PCR amplification using 16s rRNA gene s pecific 
primer set. All isolates were found to be positive for E. 
rhusiopathiae with amplification products of 407bp. 



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500 bp 
407 bp  

 
  

 
 

 

100 bp 
 
 

 
Figure 2. 2% agarose gel show ing PCR Products. Here, L1- 
100 bp marker, L2 NTC,  L3& 4 PCR products.  

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
Figure 3. Phylogenetic tree of Nagaland isolates of E. rhusiopathiae (KT160358, 
KT160359 as compared to the ATCC 19414 type strain and other isolates. 

 
 
 

(Figure 2).  
Two Nagaland isolates KT160358 and KT160359 were 

sequenced and showed high sequence identity with the 
sequences of other E. rhusiopathiae available in the 
GenBank database. The Nagaland isolates (accession 

 
 
 
 

numbers KT160358, KT160359) were in the same clad 
(Figure 3) along with other strains originated from China. 
Phylogeny based on the nucleotide and amino acid 
sequences of the viruses provides a better understanding 
of the molecular epidemiology of the isolates. The state 



5 

 

   
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
Figure 4. Histopathology of skin biopsy sample, H&E, 100X. Congestion and 
presence of microthrombi in the capillar ies of the der mis along w ith 
infiltration of lymphoid cells and fibroblast w ere consistent finding. 

 
 

 

Nagaland shares international boundaries with My anmar 
and having territorial link with other neighboring 
international countries like China, Bhutan and 
Bangladesh. The Chinese strains used in the construction 
of phylogenetic tree viz. swine isolates KPO  
63150.1, strain Er.GXBY-1(KP063149.1), strain 
Er.GXLC-1(KP063151.1) and murine isolate  
DQ4625711are forming same clad with Nagaland 
isolates ( KT160358 and KT160359) indicated an 
epidemiological link. Movements of animals and animal 
products might spread the infection to this locality. 
 

 

Histopathology 

 

Histopathologic alteration of affected skin showed 
extensive damage to the capillaries and venules of 
dermis with infiltration of lymphoid c ells and fibroblasts. 
Congestion and pres ence of microthrombi in the 
capillaries were consistent finding in all skin biopsy 
samples investigated in the present study (Figure 4). 
Similar observations were also recorded by Shankar et 
al. (2009).  

Cultural characteristics, molecular confirmation and 
histopathological changes conclusively proved that swine 
erysipelas is prevailing in Nagaland. Pigs harbouring the 
infection manifest clinically at sudden change of climate. 
It is estimated that 30–50% of healthy swine harbour the 
organism in their tonsils and other lymphoid tissues 
(Stephenson and Berman, 1978). Again, trans border 
movement of pigs and their products could facilitate 
spreading of SE in this locality. Further study on 

 
 
 

 

prevalence of swine erysipelas and identification of 
carrier pigs can provide actual guidelines to control the 
disease in this part of India. 
 
 
Conclusion 

 

Unorganized pig farms in the Phek district of Nagaland 
experienced high mortality of grower pigs during rainy-
summer season. A detail isolation and molecular 
investigation confirmed the association of swine 
erysipelas in this part of north eastern region of India. 
This is the only report on seasonal occurrence of swine 
erysipelas in the hilly low temperate climate of the NE 
states of Nagaland, India. 
 

 

Conflict of Interests 
 

The authors have not declared any conflict of interests. 

 

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