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Brucella canis

Brucella canis is a Gram-negative bacterium in the family Brucellaceae that causes brucellosis in dogs, other canids, and in rare cases, humans. It is a non-motile short-rod or coccus-shaped organism, and is oxidase, catalase, and urease positive. B. canis causes infertility in both male and female dogs. It can also cause inflammation in the eyes. The hosts of B. canis ranges from domestic animals to foxes and coyotes. It is a zoonotic organism, meaning it is able to be passed from animals to humans. It is passed from species to species via bodily fluids such as genital secretions and urine. Treatments such as spaying, neutering, and long-term antibiotics have been used to combat B. canis in dogs. The species was first described in the United States in 1966 where mass abortions of beagles were documented. Brucella canis can be found in both pets and wild animals and lasts the lifespan of the animal it has affected. B. canis has two distinct circular chromosomes that can attribute to horizontal gene transfer.

Morphology
Brucella canis are non-motile organisms and cannot move independently due to the absence of flagella. Brucella are also non-encapsulated, non-spore forming bacteria that replicate in the ER of their host cells. The bacteria are Gram-negative coccobacilli or short rods measuring 0.6 to 1.5 μm long, and 0.5 to 0.7 μm wide, do not have a capsule, do not form spores, and are aerobic. On blood or chocolate agar, colonies are small (~0.5-2 mm after 48-72 hours), convex, non-hemolytic and non-pigmented. Often B. canis colonies can present themselves as rough variants, a reflection of their naturally rough lipopolysaccharide (LPS) phenotype. The optimal growth temperature for B. canis is 37°C, but growth is still possible within the range from 20°C to 40°C. Additionally, the pH range in which B. canis grows most effectively is from pH 6.6-7.4, making this organism neutrophilic in nature. Importantly, B. canis is a "natural rough" Brucella. Its Lipopolysaccharide lacks the O-polysaccharide that is present in smooth strains; an envelope trait that influences colony phenotype and host interaction without implying a-virulence. B. canis is also unique from other Brucella species in that they demonstrate a distinctive phospholipid arrangement that differ greatly from other Gram-negative bacteria. Their envelope incorporates uncommon lipid species such as altered phosphatidylethanolamine and lipid A derivatives enriched with long-chain and branched fatty acids; features that reflect evolutionary adaptation to an intracellular lifecycle. Additionally, its phospholipid portion is mainly composed of cis-vaccenic cyclopropane with small amounts of lactobaccilic acid. This differs from other Brucella species, as they demonstrate the opposite composition, with lactobacillic acid making up the majority of the phospholipid fraction. Brucella is unusual in this composition because lactobacillic acid is typically within Gram-positive organisms but not common within Gram-negative organisms such as Brucella. These specific envelope features are discussed alongside the organisms hallmark intracellular cycle. After uptake, Brucella replicate within ER-derived Brucella containing vacuoles, a niche specific to replication and survival within B. canis. Identification B. canis is a zoonotic organism. The bacteria are oxidase, catalase and urease positive and non-motile. Unlike haemophilus, which they resemble, they have no requirements for added X (hemin) and V (nicotinamide adenine dinucleotide) factors in cultures. Full identification is established by serology and PCR. Due to B. canis being naturally rough (lacking O-polysaccharide), smooth-antigen serology is unreliable. Modern practice uses B. canis-adapated serological assays such as RSAT/2-ME, IFAT (Immunofluorescence Antibody test) and ELISA to make full identification. B. canis is not acid-fast, but they tend to maintain their color when exposed to weak acids. This results in their red color when stained. When isolated, B. canis is always in a rough form, with hydrophobic LPS imbedded in its outer membrane. MALDI-TOF mass spectrometry with validated databases and whole-genome sequencing (WGS) are now increasingly used for definitive confirmation of B. canis and specific outbreak tracing. Colonies of Brucella can typically start to be seen after 48 hours. These colonies tend to be 0.5-1.0 mm in diameter, with a convex shape and are typically circular. Growth is often slower than other bacteria, with colonies requiring up to 72 hours for clear visualization. B. canis also possesses a complete tricarboxylic acid (TCA) cycle, which primarily utilizes oxygen as its terminal electron acceptor within its electron transport chain. In anaerobic conditions, nitrate can also function as a terminal electron acceptor because B. canis is capable of producing nitrate reductase. B. canis also exhibits strong urease activity, producing the enzyme urease to hydrolyze urea into ammonia and carbon dioxide. This enzymatic activity is relevant for its role in nitrogen acquisition and as a notable virulence factor, as it helps to neutralize and facilitate survival within surrounding acidic environments. For laboratory identification, a relevant metabolic characteristic of B. canis is that it does not require supplemental for growth, unlike some other Brucella species. Additionally, B. canis has demonstrated growth on media containing thionine, but no growth on media containing basic fuchsin. Genome B. canis has two distinct circular chromosomes, a structure conserved across the Brucella genus. For the reference strain ATCC 23365, Chromosome 1 has 2,199 genes, and Chromosome 2 has 1,224 genes. These two circular chromosomes contain multiple distinct shared regions, which can be attributed to horizontal gene transfer. Despite this significant similarity, it is still possible to differentiate between B. canis and B. suis using PCR assays targeting specific known genetic variations. The most notable distinguishing factor is the lack of O-polysaccharide in its lipopolysaccharide, causing the naturally "rough" phenotype for B. canis in contrast to B. suis which retains the naturally "smooth" phenotype. ==Pathogenicity ==
Pathogenicity
The disease is characterized by epididymitis and orchitis in male dogs, endometritis, placentitis, and abortions in females, and often presents as infertility in both sexes. Other symptoms such as inflammation in the eyes and axial and appendicular skeleton; lymphadenopathy and splenomegaly, are less common. Signs of this disease are different in both genders of dogs; females that have B. canis infections face an abortion of their developed fetuses. Males face the chance of infertility, because they develop an antibody against their spermatozoa. This may be followed by inflammation of the testes which generally settles down a while after. Another symptom is the infection of the spinal plates or vertebrae, which is called diskospondylitis. It is generally spotted in the animal's reproductive organs. This infection usually causes the animal to spontaneously abort a fetus and can also cause an animal to become sterile. Host range The host range of the bacterium is mainly domestic dogs but evidence of infections in foxes and coyotes has been reported. In their urine, infected dogs can carry up to 106 bacteria per milliliter, compared to the genital discharges that can carry up to 1010 bacteria per milliliter. Neutered dogs are also capable of shedding the bacteria in their saliva and nasal secretions. Transmission to humans is rarely diagnosed but is possible. It can be transmitted via bodily fluids and aborted material. Signs and symptoms are very non-specific, such as fever, joint pain, and fatigue. Antimicrobial treatment and sterilization of the infected animals is considered an alternative to removing the animal. B. canis is mainly found in dogs, but can also affect other wild canine species such as wolves, foxes, and coyotes. The bacterium persists in these hosts, being an adaptive pathogen towards canines. The environment these hosts reside in further contribute to the canine specificity. Dog kennels are a favored environment for the bacterium to spread due to transmission through urine and reproductive fluids. Movement of infected animals through pet trade or shelter transport also plays a critical role in the broader distribution of the bacterium. In wildlife, B. canis is also circulated through hunting and scavenging. Zoonotic transmission to humans is rare but possible, particularly for people in close contact with infected reproductive tissues. == History ==
History
Brucella canis was first discovered in the United States by Leland Carmichael in 1966, when the bacterium was identified in canine vaginal discharge and the tissues from mass abortions in beagles. By the 2000s, molecular analysis revealed that B. canis is unique from other Brucella species, categorized as a naturally occurring rough species that needs an anti-rough lipopolysaccharide reagent for detection. B. canis has been recognized as a growing zoonotic risk worldwide, having the ability to infect humans. Laboratory exposure or contact with pet dogs that had aborted fetuses were the main sources of exposure to the bacterium. Human infections are often underdiagnosed due to limited diagnostic awareness and the overlapping clinical presentation with other Brucella species, which hinders precise identification. ==References==
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