**7.7 Urine analysis**

Routine test may reveal presence of bilirubin, haemoglobin and protein in the urine. In a recent study conducted to assess renal dysfunction in *Babesia rossi*, infection revealed higher concentrations of urinary IgG, urinary CRP and urinary RBP suggestive of both glomerular and tubular dysfunction [51]. These can be used for early detection of *Babesia*-induced renal dysfunction than serum urea and creatinine and urine specific gravity. There will be minimum and variable changes in urine enzyme activity (GGT and ALP) in babesiosis, thus limiting their use as diagnostic tests [47].

## **7.8 Serological diagnosis**

A recent study demonstrated the opportunity of an early and specific detection of acute infections by an AgELISA that is potentially translatable to a rapid diagnostic test design and can be used in an ELISA to detect circulating *Babesia* antigen during acute infections and can be used to detect parasites 24–48 h before it could be detected by light microscopy [52].

#### **7.9 Molecular diagnosis**

Molecular techniques help in refining the diagnosis to the species level and thus provide a more accurate prognosis. Different molecular techniques are used for identification and differentiation of the various species of *Babesia*, semi-nested PCR [53] reverse line blotting [9, 54] and PCR-restriction fragment length polymorphism analysis [55].

Improved PCR techniques have lately allowed for better definition of these parasites [56] and allows for a more reliable identification of the etiological agents compared to direct detection by light microscopy or serology [2]. It is more sensitive and provides an evidence of an active and ongoing infection in a clinical setting.

In addition, several genes are commonly used to discriminate among *Babesia* species. Typically, these include the nuclear ribosomal RNA genes [7, 8] and the two internal transcribed spacers (ITS1 and ITS2) [7]. PCR DNA amplification can be a useful technique for monitoring treatment [57].

## **7.10 Diagnostic imaging**

The most consistent findings on abdominal ultrasonography in dogs with *B. canis* are hepatomegaly and splenomegaly with diffuse, hypoechoic, heterogeneity and renal changes like diffuse homogenous increased cortical echogenicity and increased corticomedullary definition [58]. Dogs infected with *B. rossi* and *B. canis* having gastrointestinal signs and abdominal pain are reported to have

ultrasonographic changes in the pancreases [32, 59] which are found to be consistent with acute pancreatitis and included duodenal atony and peripancreatic fat hyperechogenicity.
