THE EFFICACY OF TM-700

FOR FISH IN MEDICATED FEED

HEALTHY ANIMALS. HEALTHY FOOD. HEALTHY WORLD.

The efficacy of TM-700 for fish in medicated feed to control mortality

caused by streptococcus agalactiae in hybrid tilapia
(oreochromis niloticus x o. Aureus)
Natan Wajsbrot (M.Sc.), Yechiam Shapira (M.Sc.), Allan Heres (Ph.D.), Yoav Rosen (M.B.A.) and
Raanan Ariav (D.V.M.)
Phibro Aqua, Israel
Case study
Introduction
Israel is located in a semi-arid zone, characterized by cold winter and dry summer, and an average annual
rainfall of around 500 mm/year, most of it occurring in the Central-North areas of the country. The only large
fresh water body is the Lake of Galilee, which mainly supplies water for human consumption. The Northo
East valleys of Israel (Harod and Beit Shean) are characterized by hot summer (up to 45 C) and cold winter
(down to 10oC). The major fish farmers in Israel are found in these regions. In spite of the climatic limits and
overall shortage of water, both agriculture and aquaculture are highly developed in Israel.
The main fish culture method is in reservoirs, which are used to store rainwater during the winter. Israeli
agriculture is today largely intensive and depends on irrigation from these reservoirs during the dry summer.
Recently, it has become common to use irrigation reservoirs for fish culture in integrated farming systems.
These integrated agriculture-aquaculture systems use the water twice: within an aquaculture production
system, and subsequently supply to irrigated agriculture systems. This system, now a few decades old, was
a significant step in the intensification of inland fish culture in Israel.
The high temperatures of the water during the summer, combined with the intensification and the massive fish
handling leads to various typical bacterial diseases due to high temperatures, like Streptococcosis disease.
Streptococcosis is an important disease that impedes development and progression of aquaculture worldwide
due to its severity and frequency in freshwater and marine farmed fish (Toranzo et al., 2005; Agnew &
Barnes, 2007). Streptococcal infection causes high mortality (up to 50 - 75%) in susceptible fish species,
particularly in intensive cultured systems (Evans et al., 2002). The disease leads to significant economic
losses in several finfish industries, including Tilapia, which is one of the most susceptible species (Chang
& Plumb, 1996). Several reports have described natural outbreaks of streptococcal infection in Nile Tilapia
(Oreochromis niloticus), red Tilapia (Oreochromis sp.) and hybrid Tilapia (O. niloticus Oreochromis aureus)
in different areas of the world, including Israel (Eldar et al., 1994).
Streptococcosis is a septicemic disease caused by gram-positive cocci bacteria of the family Streptococcaceae.
The main etiological agents of warm water streptococcosis in fish consist of Streptococcus agalactiae,
Streptococcus iniae, Streptococcus dysgalactiae (Nomoto et al., 2004; Yang & Li, 2009) and Lactococcus
garvieae (Kusuda et al., 1991). However, the major pathogens reported in Tilapia are S. agalactiae and S.
iniae (Eldar et al., 1994; Shoemaker et al., 2001).
Due to the recent commercial growth of Tilapia culture, streptococcal infections are becoming a major threat
to the Tilapia industry. The disease leads to morbidity and mortality with acute or chronic infection. Moreover,
subclinical infection reduces fish appetite, which consequently increases the duration of grow-out. Also, the
physical appearance of the infected fish, such as missing eyes or hemorrhagic skin, and the poor meat
quality are not acceptable to the consumer. The infected fish are also injured easily during harvesting and
transportation, which leads to a shorter shelf life of the product.

The streptococcal pathogen responsible for disease outbreaks was characterized using conventional
bacteriology, gram stain and molecular identification to confirm the spread of disease in Tilapia farm naturally
infected by these bacteria.
Case study description
Date: August, 2014
Place: Beit Shean Valley, Israel
Fish pond type: Reservoir (Fig. 1).
Reservoir size: 13 hectares. Average depth: 3 meters.
Stocking Density: 400,000 Tilapia (420 g average weight) and 50,000 Flathead grey mullet (Mugil cephalus)
(380 g average weight).
History: During the summer in August 2014 dead fish were observed around the reservoir. Figure 2 shows
the cumulative mortality of fish in the first two weeks of the outbreak. Weak fish were collected for pathological
examination in Phibro Aqua field diagnostic laboratory located in Kibbutz Kfar Ruppin (Beit Shean Valley).
Water quality parameters:
-
-
-
-
-
-

Temperature: 28C
Dissolved oxygen: 3.1-5.9 ppm
Ammonium: 1.0 mg/l
Nitrite: 0.6 mg/l
Total salinity: 1.5 ppt
pH: 7.5

Figure 1: Earthen reservoir in Beit Shean valley. The size of the reservoir is 13 hectares and the average
depth is 3 meters. It can grow up to 200 tons of fish from different species.

Figure 2: Cumulative mortality of fish along the reservoir edge. This mortality was due to a Streptococcosis
outbreak in the summer of 2014. The average weight of the fish was 380-400 g, showing typical symptoms
of the disease.

Results of the fish examination

Parasitology:
Skin: Trichodina spp., Sessiline spp., and Gyrodactylus spp.
Gills: Trichodina spp. and Sessiline spp.
Internal organs:
Spleen: MMC (Melano Macrophage Center) at high level.
Septicemia and internal bleeding.
Gross pathology: Exophthalmia was observed in symptomatic fish (Fig. 5).

Gram stain
Figure 3: Gram-positive cocci from Tilapia kidney smear (X100).

Bacteriology results
Fish brain and kidney tissues were taken for bacteriological identification and antibiogram assays (disk
diffusion method).
The bacteria was isolated and culture on a Blood TSA agar plate and sent to Hy-Labs Laboratories LTD for
molecular identification. The laboratory confirmed its identification as Streptococcus agalactiae.
Figure 4: Antibiogram on Blood TSA agar. In this disk diffusion method the size of the circular inhibition
zone demonstrated the sensitivity of the bacteria to the antibiotic. The disk (BD-BBLTM) contained 30 g of
the active ingredient in TM-700 and showed antimicrobial efficacy against S. agalactiae.

Antibiogram result
Antibiotic

Disc potency (g)

Zone of Inhibition
(diameter in mm)

Active ingredient in
TM-700

Antibiotic Sensitivity
Profile

30

Sensitive

23

Figure 5: Typical clinical sign in Tilapia associated with Streptococcosis is exophthalmia.

Once the antibiogram assay was completed the fish were treated with TM-700 at a dose of 100 mg AI (active
ingredient)/Kg of BW (body weight)/day for 14 days.
After 5 days of treatment fish morbidity and mortality significantly decreased until the complete cessation of
mortality at day 14 of treatment.

Conclusion:
In the current Streptococcosis outbreak in Tilapia, TM-700 proved to be very effective against Streptococcus
agalactiae after 14 days of treatment with medicated feed.
Reference
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