**3. Some examples for the effect of probiotics in helminth parasitic zoonosis**

#### **3.1 Effect of probiotic on Cryptosporidium**

Cryptosporidium is an abdominal pathogen followed by the Alveolata group. It could cause overwhelming contamination of gastrointestinal in immunosuppressed humans. In the surroundings, the infective form Cryptosporidium is determined as oocyst found in water. After consumption, the oocysts passage through the gut lumen to the small intestine, in which they release the motile sporozoites that adhere and attack the epithelial intestine cells. The sporozoites are disrupting the microvilli and penetrate to the cells of the host to arrange their intracellular position, wherein they continue to be in a further cytoplasmic vacuole. After replication of parasite and elusion, oocysts are produced and excreted within the faces [36, 37]. Intestinal epithelial cells, inflamed by way of *Cryptosporidium parvum*, show lessened sodium ions and water absorption as well as greater chloride ion secretion, leading to diarrhea. Also, the use of paromomycin and azithromycin or nitazoxanide is the handiest powerful in mixture with immune restoring agents [38]. Beneficial outcomes of probiotics upon cryptosporidiosis have been tested—female mice were fed day-by-day for 4 weeks vintage with *L. reuteri* traces 4000 and 4020 or *L. acidophilus* NCFM provided reduced oocyst dropping [39, 40]. Waters et al. [41] recommended that protection becomes due to secretion of as yet unidentified antimicrobial products. Curiously, *in vitro* research tested the inhibitory results of cellular-unfastened supernatants of *L. acidophilus* NCFM and *L. reuteri* stress 23,272 on *C. parvum* and *C. hominis* viability and infectivity [42, 43]. In addition, mobile-unfastened supernatants of *Bacillus brevis*, *Enterococcus faecium,* and *Pseudomonas alcaligenes* reduce *C. parvum* oocyst persistence by inducing oocyst premature excystation [44, 45]. The compounds at the basis of such inhibition are under investigation. Pickerd and Tuthill, [46] resolved diarrhea due to *Cryptosporidium oocysts* via probiotic lines *Lactobacillus rhamnosus* GG 109 CFU/day and *Lactobacillus casei* Shirota 6.56 X 109 CFU/day during 4-week. Within 10 days of beginning, the pain was decreased and the pattern of stool unfastened from Cryptosporidium after 4 weeks after beginning with probiotics.

Also, Sanad et al. [47] studied the therapeutic effects of daily administration of a mixture of *L. acidophilus, L. helveticus,* and *Bifidobacterium bifidum against C. parvum* infection in an immunosuppressed mouse model. The parasite was not achieved by eradication. But, the infection of the probiotics used caused a significant reduction in parasite burden, ultrastructural changes with respect to parasite attachment, internalization into epithelial cells, partial compensation of the mucosal damage caused

by the parasite, and an increase in serum level IFN. These results reveal the beneficial effects of probiotics on cryptosporidiosis and suggest that they can help to reduce the risk of serious disease in immunosuppressed patients.

Moreover, Del Coco et al. [48] studied the treated *C. parvum* infection by oral administration of *Enterococcus faecalis* CECT 7121 as probiotic strain in immunosuppressed mice. Also, studied the effect of *C. parvum* infection in the intestinal mucosa and counted at each part of the intestine. The results were established that when both *E. faecalis* and *C. parvum* were present in the same intestinal location happened interfered with them. Also, proposing that supplementation of *E. faecalis* can improve the harmful effects on infection of *C. parvum*. Also, Glass et al. [42] recognized that *L. acidophilus* (LA) and *L. reuteri* (LR) cell-free supernatants able to diminish about 21–42% and 30–35%, respectively of the infection of bovine *C. parvum* and *C. hominis* in a cell-culture immunofluorescence (CCIF) test. Moreover, reduction of oocyst viability reached 40–80% at 24 h incubation of bovine *C. parvum* oocysts in the bacterial cell-free supernatants and this reduction was evaluated by flow cytometric analysis and the infectivity of oocyst reached up to 95% by the CCIF analysis. So, the production of antimicrobial compounds secreted from LA and LR had a harmful effect on bovine *C. parvum* and *C. hominis*. Likewise, Khalifa [49] evaluated a study consisting of 70 mice as; 60 mice were infected with *Cryptosporidium oocysts and* immune-suppressed, other 10 mice were not infected and left immune-competent. Formerly, the mice were divided into three groups; group (1) infected mice were treated with *L. casei*, group (2) infected mice were treated with yogurt, and group (3) infected mice but not treated as control. The counts of oocyst in the mice stools were determined to evaluate the cryptosporidiosis progress and measured by the developmental stages in histopathological sections of ilea. The results found that the parasitic burden in mice was reduced by regular administration of yogurt and *L. casei* in comparison with the control group. Moreover, the use of yogurt daily was more effective than *L. casei* where the yogurt was stopped oocyst shedding previous than *L. casei* and the counts of oocyst were lesser during the experiment duration in comparison with infected mice that treated with *L. casei*. Previous studies indicated that the used probiotics are promising and hopeful to control and treated the parasite's development.

In contracts, Guitard et al. [50] studied the feeding rates with *L. casei* daily with 2 × 107 CFU before 2 days of the infection until the spontaneous clearance of the parasite. Effects on weight gain, parasite burden, mucosal histology, and production of mucosal cytokines (IFNγ, IL10, and TNFα). The authors also indicated that administration of probiotic strains through the infection course was not significantly affected the weight gain, parasite burden, mucosal damage, or mucosal cytokines kinetics. Overall, the studied model data revealed that the use of *L. casei* as regular administration was unable to eradicate the parasite. Other studies established that treatment the cryptosporidiosis by probiotic strains did not eradicate the parasite, but resulted in a moderate benefit with a decrease in parasite burden and mucosal damage, and these results were obtained after long-term feeding (7–28 days) or prolonged pre-feeding (≥7 days) before infection [51–55].

#### **3.2 Effect of probiotics on Giardia**

*Giardia lamblia* (also known as *Giardia intestinalis* or *Giardia duodenalis*) is an intestinal pathogenic protozoan parasite belonging to the Diplomonad institution that reasons ∼280 million symptomatic human infections in line with 12 months [56]. This

#### *Perspective Chapter: Application of Probiotics to Inactivate Helminth Parasitic Zoonosis DOI: http://dx.doi.org/10.5772/intechopen.103744*

monoxenous waterborne parasite has the capability to contaminate an extensive variety of hosts. To initiate the infection for humans need, 10 environmentally resistant cysts to infect. When the cysts passage through the gastrointestinal, they unlock and replicates to form trophozoites. These trophozoites had the ability to reproduce inside the gut lumen and adhere to the epithelium. These proliferate of trophozoites in the gut was associated with the disorder symptoms, such as watery diarrhea, epigastric pain, nausea, vomiting, and weight drop, which appeared during 6–15 days after cyst consumption, but half of the infections stay asymptomatic. The infection was mainly treated by metronidazole and nitroimidazole, but infections can also solve spontaneously. The immune response T cells, neutrophils, macrophages also with IgM, IgG, and IgA antibodies are major players for the decision of giardiasis. *L. casei* MTCC 1423 stress as well as *E. faecium* SF68 were additionally effective in eliminating Giardia contamination from mice [53, 57]. Protection becomes related to a diminution of atrophied villi and infiltrating cells inside the small gut of probiotic-handled mice [57] or with an enhancement of the immune response for the reason that production of specific anti-Giardia intestinal IgA and IgG was noticed in dealing with mice. *In vivo* experimentation on malnourished mice showed that day-by-day pretreatment with *L. casei* MTCC1423 effectively decreased severity and period of giardiasis, as compared to non-probiotic-fed malnourished mice [58, 59].

Shukla et al. [60] determined the acid-tolerated strains of probiotics *L. casei* or *Lactobacillus* yogurt when found in the gastrointestinal tract. The authors have studied the possibility of these isolates to therapeutic treated the infected mice with the Giardia. After 1 day of Giardia infection, it was found that supplementation of probiotics either *L. casei* or L. yogurt were eliminated the infection severity comparison with Giardia infected mice. All changes in the Pathophysiologically, the morphological and cellular changes of the small intestine were slightest in treated mice with probiotics in compared to harshly inflamed, edematous, vacuolated epithelial cells in infected mice with Giardia. The results concluded that L. yogurt possessed better probiotic properties and has the possibility to diminish the severity of infection in mice with Giardia. Also, Goyal et al. [61] investigated the efficiency of four probiotic strains (*L. rhamnosus* GG (LGG), *L. acidophilus*, *L. plantarum,* and *L. casei*) against the murine giardiasis modulation. The daily strain was received around 109 CFU for single animal via orogastric gavage. The more effective strain was LGG, which proved more effect in decreasing the duration of *G. lamblia* cycle, by eliminating the active trophozoite number in the intestine, increasing cyst excretion, and leading to suppression of the disease around 13 days after trial inoculation. Amer et al. [62] evaluated *in vitro* and *in vivo* the beneficial effectiveness of bacteriocins that resulted from new Egyptian probiotic Lactobacilli strains [*L. acidophilus* (P106) and *L. plantarum* (P164)] against *G. lamblia*. The results showed that 50 μg of bacteriocin from *L. acidophilus* eliminated the trophozoites adherence and the counts around 58.3 ± 4.04%. Oral feeding of 50 μg/mouse of bacteriocin from *L. acidophilus* every 5 days was able to reduce the density of parasites inside the intestines and enhance the strength of the gut disease system of infected mice. The authors established that bacteriocin from strain *L. acidophilus* (P106) had a promising potential therapeutic outcome and alternative safety way instead of present commercial drugs to treat *G. lamblia*.

In the same line, the Bifidobacterium efficiency can be evaluated in an experiment against infected mice with *G. lamblia* infection. The single-dose about 0.1 ml of Bifidobacterium cells for every day significantly eliminated the *G. lamblia* cysts shedding in feces, and this infection was disappeared totally at the 5 days of probiotic Bifidobacterium inoculation. Also, in the mice group that used metronidazole, the

authors found that Giardia cysts were reduced and infection cured on the day 17th of treatment, in comparison with the control group that showed parasite shedding cysts. Moreover, for histopathological results, *in vivo* by gut cells, the Bifidobacterium has prevented inflection of the Giardia colonization and able to reduce the infection with this parasite [63]. Generally, the usage of probiotic strains, such as *Lactobacillus* and *Saccharomyces*, had a positive influence to reduce gastrointestinal symptoms time and repair the damages, especially for giardiasis. Probiotics had the ability to control the composition of commensal microbiota and balance which lead to therapeutic impact. According to pre-clinical and clinical searches, different probiotic strains can increase the antioxidant capacity, destroy oxidative products, regulate the systemic, and activate the responses of mucosal immune as well as reduce gastrointestinal symptoms time, that lead to protect against mucosal damages that induced by parasites. In addition, they can reduce the *G. duodenalis* proportion load by directly targeting the parasite. They can produce some anti-giardial factors that feature destroy the parasite's cellular architecture and suppress the parasite's proliferation and growth [64, 65].

### **3.3 Effect of probiotics on Eimeria**

Eimeria is an apicomplexan parasite responsible for formed coccidiosis found in poultry, cattle, rabbits, dogs, and cats mainly small animals. A primary parasitic ailment in poultry is avian coccidiosis, with a major economic importance worldwide impact [66]. When the chickens ingest oocysts, it is become inflamed and eventually exocyst to shape sporozoites within the lumen of the top gut. Those sporozoites migrate to their preferred sites of development. They then invade villi enterocytes and undergo a first asexual multiplication, the schizogony, leading to the discharge of numerous merozoites that initiate a 2d schizogony via infecting new epithelial cells. Macro- and microgametes are finally produced, starting up the sexual phase that yields environmental resistant oocysts which might be shed within the feces [67]. Two main ways to control this parasite are by drugs, such as amprolium, halofuginone, and monensin lasalocid, or live vaccines. But stay vaccines against coccidiosis are incredibly effective, primarily based on non-attenuated and attenuated lines.

Probiotic supplementation can enhance performance and help alleviate the negative effects of a mixed *Eimeria* infection. The study by Ritzi et al. [68] evaluated the effects of probiotics on birds and resistance to a mixed *Eimeria* infection in commercial broilers. Using treatments, including negative control (non-infected, NEG), positive control (*Eimeria*-infected, PoS), anticoccidial control (0.01% salinomycin, SAL), irregular high-dose water-applied probiotic (WPI), irregular low-dose water-applied probiotic (WPC), and feed-supplemented probiotic (FSP). On day 15 of the experiment, all birds except those in NEG were treated with a mixed dose of *Eimeria acervulina*, *Eimeria maxima*, and *Eimeria tenella*. Samples of birds feces were collected from day 20 to 24 for counts of the oocyst and evaluated the lesion scores were at day 21. The probiotic groups were comparable with the birds for SAL-treated, except during the 6 days immediately following the *Eimeria* species challenge, where the SAL birds displayed well performance. The lower duodenal and jejunal lesion scores were found for WPC birds, signifying a healthier intestine and improving the resistance to *Eimeria* species in comparison with the positive control (PoS). Also, fewer oocysts in the feces were recorded for birds in the WPI treatment, although this was not a trend for all of the probiotic treatment groups. The addition of probiotic secretion compounds containing *Pediococcus acidilactici* in the ration of the birds before experimental infection with *E*. *tenella* resulted in mild improvement in the performance parameters, a slight reduction

*Perspective Chapter: Application of Probiotics to Inactivate Helminth Parasitic Zoonosis DOI: http://dx.doi.org/10.5772/intechopen.103744*

in lesion score, and in the oocysts count when compared with the birds treated with anticoccidial drugs, but that picture was better than the infected non-treated group. The addition of compounds containing natural microflora (especially those producing lactic acid) to the poultry feed or water to overcome coccidial infection especially [69]. Also, chickens fed on *Lactobacillus*-based ration showed reduced oocysts output compared to controls after challenge with *E*. *acervulina* [70].

**Another effect of probiotics was observed on malaria** as the recent study by Elli et al. [71] investigated the use of probiotic *L. casei* in treating malaria in mice with chloroquine. Probiotics in combination with chloroquine showed complete suppression in the parasitemia rate. The data were established by histological observation of two major organs, the liver and spleen. Interestingly, further suppression of parasitemia and hemosiderosis was observed when probiotic was given along with chloroquine. Another author Oliveira-Sequeira et al. [72] have shown a reduction in the number of *Strongyloides venezuelensis* in infected mice about 33% and egg output upon feeding with probiotic *Bifidobacterium animalis*, and probiotics was improved the immune responses. A new study has linked the microbiome of the human gut with immunity against malaria infections. Gut probiotics represent innovative tools for malaria prevention and lead the way to novel types of vaccination strategies [73, 74].
